A freshwater fish medicated bath traditional Chinese medicine preparation, a preparation method and application thereof

Abstract

A traditional Chinese medicine preparation for freshwater fish bath therapy, its preparation method, and its application, relating to the field of aquatic drug technology, includes the following steps: S1, adding Bletilla striata extract and Ulmus pumila bark extract to water to obtain solution I; S2, dissolving Isatis indigotica leaf extract, Salvia miltiorrhiza extract, Paeonia suffruticosa bark extract, Scutellaria baicalensis extract, Phellodendron chinense bark extract, Coptis chinensis extract, Angelica dahurica extract, Gleditsia sinensis thorn extract, and Pseudolarix amabilis bark extract in ethanol solution to obtain solution II; S3, mixing solution I and solution II, homogenizing under high pressure, and spray drying to obtain microcapsule drug A; S4, mixing rhubarb extract, Sophora flavescens extract, and Galla chinensis extract to obtain drug B; S5, mixing drug A and drug B to obtain the freshwater fish bath therapy preparation. The bath therapy preparation obtained by this invention can solve the problem that current freshwater fish bath therapy preparations do not provide continuous protection for fish after bathing, even when the fish are introduced into the aquaculture water.

Description

Technical Field

[0001] This invention relates to the field of aquatic drug technology, specifically to a traditional Chinese medicine preparation for freshwater fish baths, its preparation method, and its application. Background Technology

[0002] Freshwater fish inevitably suffer injuries such as scale loss and damage to their skin during netting, pond transfer, and transportation due to friction, squeezing, bumping, and stress. Fish live in water and are normally not prone to disease. Their scales, mucus, and skin form their first line of defense. Wounds disrupt this barrier function, leading to bacterial and fungal infections and various fish diseases, causing significant economic losses to aquaculture. Currently, external chemical disinfectants (such as trichloroisocyanuric acid, hypochlorous acid, chlorine dioxide, and potassium permanganate), bactericides (such as antibiotics), and natural medicines (including traditional Chinese medicine, medicinal plants, and their extracts) are mainly used to prevent and treat fish injuries and various secondary diseases. Treatment methods mainly include medicated baths and full-pond spraying. Medicated baths effectively kill pathogens on the fish's body surface, repair wounds, and prevent and treat various fish diseases caused by injuries. They have advantages such as relatively low dosage, simple and convenient operation, and easily controllable risks. However, after a medicated bath, the amount of medication on the fish's body surface becomes negligible after dilution, failing to provide continuous protection and leaving them still at risk of wound infection and pathogen invasion. Full-pond spraying disinfects and sterilizes the entire aquaculture water body, killing most pathogens and providing continuous protection for the fish. However, it also has disadvantages such as high dosage, potential water quality degradation, high risk, and difficulty in controlling the dosage.

[0003] In view of the above problems, it is necessary to invent a fish medicine that has the advantages of both medicated baths and sprays, while effectively avoiding the disadvantages of both, and provides a broad-spectrum bactericidal and antifungal aquatic fish medicine with preventive and therapeutic effects on a variety of common fish diseases and fungal diseases.

[0004] Traditional Chinese medicine (TCM) has advantages such as fewer side effects, less irritation, green safety, and a broad antibacterial spectrum, overcoming to some extent the drawbacks of chemical disinfectants and antibiotic bactericides. For example, patent application number 201711432977.0 discloses a pure TCM disinfectant, which has the advantages of using harmless, green, and safe raw materials that are harmless to fish and humans, and has preventive and therapeutic effects on water mold disease in bass. However, as a whole-pond application, its dosage reaches 50-120g / kg of water, which has the disadvantages of large dosage and high cost, limiting its applicability.

[0005] Patent 200910210511.5 discloses a pure Chinese medicine disinfectant and medicated bath preparation, which has the advantages of harmless, green and safe raw materials. It has the effect of preventing and treating perch trichodiniasis, but no significant effect on fish injuries and bacterial diseases.

[0006] A traditional Chinese medicine preparation for treating hemorrhagic disease in silver carp (201510933445.X) involves decocting herbs such as Astragalus membranaceus, Bupleurum chinense, and Fritillaria cirrhosa, then diluting the decoction and using it to bathe silver carp. This can effectively prevent and treat hemorrhagic disease in silver carp. However, after the fish enter the aquaculture water, the medicine cannot provide continuous protection for the fish.

[0007] The patented cefadroxil and Bletilla striata capsule for livestock and poultry (application number 200510122494.1) mixes cefadroxil and Bletilla striata and then puts it into a capsule. It can prolong the retention time of the drug in the lesion and increase the local concentration of the drug in the diseased tissue. It has good bioadhesion, but it is not suitable for use in aquatic animals.

[0008] A patent for a borneol camphor essential oil nasal spray and its preparation method (application number: 202110186780.3) describes a method of filling borneol camphor essential oil into Bletilla striata polysaccharide microcapsules. The nasal spray can effectively inhibit bacteria and cleanse the skin, and is suitable for daily health care of allergic rhinitis and nasal discomfort such as nasal congestion, nasal itching, and impaired nasal mucosa function. After adopting microcapsule technology, the borneol camphor essential oil is solubilized and enters the aqueous system, which plays a role in the sustained release of natural borneol in the borneol camphor essential oil. Since fish are always in an aquatic environment, and their body surface has scales and mucus, the solubilization properties are not suitable for fish medication. Instead, it is necessary to strengthen the adhesion, sustained release, and breathability properties to reduce the amount of drug entering the water. Therefore, it is not suitable for fish medication.

[0009] A patent for an antiviral Chinese medicine sustained-release capsule for aquaculture and its preparation method (application number: 202210528295.4) discloses an antiviral Chinese medicine sustained-release capsule comprising a Chinese medicine core material and a wall material covering the surface of the core material. This antiviral Chinese medicine sustained-release capsule can simultaneously prevent and treat viral diseases in fish in both deep and shallow water areas, with good antiviral effects and minimal impact on water quality. However, this antiviral Chinese medicine sustained-release capsule mainly exerts its effects after being ingested by fish. The Daqingye and Yuhucao in the wall material, while playing an antiviral role, also act as feeding attractants, ensuring that the antiviral Chinese medicine sustained-release capsule is successfully ingested by fish. It is not suitable for external use in fish baths. Summary of the Invention

[0010] The present invention aims to provide a traditional Chinese medicine preparation for freshwater fish baths, its preparation method and application, in order to solve the shortcomings of current freshwater fish bath preparations that cannot provide continuous protection for fish after they enter the aquaculture water.

[0011] To solve the above technical problems, the specific solution adopted by the present invention is as follows: a method for preparing a traditional Chinese medicine preparation for freshwater fish baths, comprising the following steps:

[0012] S1, add Bletilla striata extract and elm bark extract to water, stir, and obtain solution I for later use;

[0013] S2, Dissolve the extracts of Isatis indigotica leaf, Salvia miltiorrhiza, Paeonia suffruticosa root bark, Scutellaria baicalensis root, Phellodendron chinense bark, Coptis chinensis root, Angelica dahurica root, Gleditsia sinensis thorns, and Pseudolarix amabilis bark in an ethanol solution, homogenize under high pressure to obtain solution II;

[0014] S3, heat solution I to 50-60℃, add solution II while stirring, keep stirring at 50-60℃ for 20-40 minutes, then homogenize under high pressure to obtain solution III, spray dry solution III to obtain microcapsule drug A;

[0015] S4, rhubarb extract, sophora flavescens extract and gallnut extract are mixed to prepare drug B;

[0016] S5. Mix medicine A and medicine B to obtain a traditional Chinese medicine preparation for freshwater fish bath.

[0017] As a further optimization of the above technical solution, the following steps are specifically included:

[0018] S1. Mix Bletilla striata extract and elm bark extract at a weight ratio of 20-30:1, add water, stir, and obtain solution I with a concentration of 5-10% for later use;

[0019] S2, according to the following weight proportions, take 200-300 parts of Isatis indigotica leaf extract, 150-200 parts of Salvia miltiorrhiza extract, 70-100 parts of Paeonia suffruticosa root bark extract, 150-200 parts of Scutellaria baicalensis extract, 70-100 parts of Phellodendron chinense bark extract, 70-100 parts of Coptis chinensis extract, 70-100 parts of Angelica dahurica extract, 80-120 parts of Gleditsia sinensis thorn extract and 80-120 parts of Pseudolarix amabilis bark extract, mix them into extract powder, dissolve the obtained extract powder in a 70-80% ethanol solution at a weight-volume ratio (g:ml) of 1:2-3, homogenize under high pressure to obtain solution II;

[0020] S3. Take 2500-3500 parts of solution I according to volume fractions, heat to 50-60℃, add 100 parts of solution II while stirring, continue stirring at 50-60℃ for 20-40 minutes, homogenize under high pressure to obtain solution III, and spray dry solution III to obtain microcapsule drug A.

[0021] S4. Take 300-500 parts of rhubarb powder, 100-150 parts of sophora flavescens powder and 100-150 parts of gallnut powder according to the weight ratio, extract, filter to remove the residue, concentrate the filtrate, dry and pulverize to obtain drug B.

[0022] S5. Mix medicine A and medicine B in a weight ratio of 1:3-5 to obtain a traditional Chinese medicine preparation for freshwater fish bath.

[0023] As a further optimization of the above technical solution, the pressure of high-pressure homogenization in step S2 is 20MPa-30MPa, and the homogenization time is 5-10min; the pressure of high-pressure homogenization in step S3 is 20MPa-30MPa, and the homogenization time is 30-40min.

[0024] As a further optimization of the above technical solution, in step S3, the inlet temperature of the spray dryer is 160-180℃ and the outlet temperature is 80-95℃.

[0025] As a further optimization of the above technical solution, step S4 is as follows: Rhubarb, Sophora flavescens and Galla chinensis are dried in an oven at 70-80℃, pulverized, and 300-500 parts by weight of rhubarb powder, 100-150 parts by weight of Sophora flavescens powder and 100-150 parts by weight of Galla chinensis powder are placed in an extraction tank equipped with a stirring device. Water is added at a weight-to-volume ratio (g:ml) of 1:15, heated, and the temperature is controlled at 70-80℃ for extraction. After filtration, the filtrate is concentrated, dried, and pulverized to obtain drug B, which is a mixture of rhubarb extract, Sophora flavescens extract and Galla chinensis extract.

[0026] As a further optimization of the above technical solution, the preparation method of the Bletilla striata extract is as follows: the Bletilla striata tuber is dried in an oven at 70-80℃, pulverized, extracted with water, filtered, and the filtrate is concentrated to obtain an extract. Anhydrous ethanol is then added to the extract, precipitated, centrifuged, the precipitate is separated, dried, and pulverized to obtain the Bletilla striata extract.

[0027] As a further optimization of the above technical solution, the preparation method of the elm bark extract is as follows: take elm bark, dry it in an oven at 70-80℃, pulverize it, add water to extract it, filter it, concentrate the filtrate to obtain an extract, introduce the extract into 95% ethanol, precipitate it with alcohol, centrifuge it, separate the precipitate, dry it, pulverize it, and obtain the elm bark extract.

[0028] As a further optimization of the above technical solution, the preparation method of the Isatis indigotica extract, Salvia miltiorrhiza extract and Paeonia suffruticosa extract is as follows: the Isatis indigotica powder, Salvia miltiorrhiza powder, Paeonia suffruticosa powder and anhydrous ethanol solution are mixed respectively, ultrasonically extracted, filtered, the filtrate is concentrated, freeze-dried and pulverized to obtain the Isatis indigotica extract, Paeonia suffruticosa extract and Salvia miltiorrhiza extract.

[0029] The preparation methods of the Scutellaria baicalensis extract, Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract, and Gleditsia sinensis thorn extract are as follows: Scutellaria baicalensis powder, Phellodendron chinense powder, Coptis chinensis powder, Angelica dahurica powder, Gleditsia sinensis thorn powder and 50% ethanol solution are mixed separately, ultrasonically extracted, filtered, the filtrate is concentrated, freeze-dried, and pulverized to obtain Scutellaria baicalensis extract, Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract and Gleditsia sinensis thorn extract;

[0030] The preparation method of the Pseudolarix amabilis bark extract is as follows: mix Pseudolarix amabilis bark powder and 75% ethanol solution, extract by ultrasonication, filter, concentrate the filtrate, freeze dry, and pulverize to obtain Pseudolarix amabilis bark extract.

[0031] A traditional Chinese medicine preparation for freshwater fish baths prepared by the method described above.

[0032] Application of a traditional Chinese medicine preparation for freshwater fish bath in the preparation of drugs for the prevention and treatment of bacterial and fungal diseases in freshwater fish.

[0033] This invention provides a freshwater fish herbal bath preparation and its preparation method. The main functions of the herbal medicines used in the herbal bath preparation are as follows:

[0034] (1) Bletilla striata: It has astringent and hemostatic properties, reduces swelling and relieves pain. The Bletilla striata gum it contains is a good polysaccharide adhesive and can be used as a drug carrier.

[0035] (2) Elm bark extract: mainly composed of glycoprotein and polysaccharide. Elm bark has excellent hemostatic effect, diuretic effect, expectorant effect, swelling reduction and detoxification effect. Adding an appropriate amount of elm bark extract to Bletilla striata can adjust the viscosity, suspension and emulsifying power of the solution.

[0036] (3) Gleditsia sinensis thorns: astringent and hemostatic, swelling-reducing and detoxifying, pus-draining, insecticidal and bactericidal effects;

[0037] (4) Moutan bark: Clears heat and cools blood, promotes blood circulation and removes blood stasis, promotes wound healing, and kills bacteria;

[0038] (5) Gallnut: It has astringent properties and inhibits water mold infection. It has an inhibitory effect on both Gram-positive and Gram-negative bacteria. It can be used to prevent and treat diseases such as white head and white mouth, myxobacterial disease, aeromonas disease, pseudomonas disease, white skin disease, red skin disease, and furunculosis in fish.

[0039] (6) Scutellaria baicalensis: antibacterial and inhibits water mold; it has inhibitory and killing effects on Pseudomonas fluorescens, Aeromonas punctata, Aeromonas hydrophila, Flexibrio columnarus, Proteus vulgaris and Edwardsiella tarda.

[0040] (7) Rhubarb: antibacterial and inhibits water mold; strong antibacterial effect, broad antibacterial spectrum, astringent, laxative, platelet-increasing, blood coagulation-promoting effects, used for gill rot, hemorrhagic disease and white head and white mouth disease, etc.

[0041] (8) Sophora flavescens: antibacterial and inhibits water mold;

[0042] (9) Phellodendron bark: replenishes qi and strengthens the exterior, astringes sores and promotes tissue regeneration, clears heat and dries dampness, clears and drains kidney fire, and kills bacteria;

[0043] (10) Daqingye: Promotes wound healing and has inhibitory and killing effects on common pathogens such as Aeromonas hydrophila, Vibrio harveyi, Aeromonas vulgaris, Vibrio anguillaensis, Aeromonas schubertii, Aeromonas enterica, and Pseudomonas fluorescens.

[0044] (11) Coptis chinensis: clears heat and dries dampness, clears the heart and drains fire, clears heat and detoxifies, and promotes wound healing; it has antibacterial, anti-inflammatory and detoxifying functions.

[0045] (12) Salvia miltiorrhiza: promotes wound healing;

[0046] (13) Angelica dahurica: astringent and hemostatic, reduces swelling and promotes tissue regeneration;

[0047] (14) Pseudolarix bark: inhibits water mold, clears heat and promotes diuresis, kills insects and relieves itching.

[0048] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0049] This invention discloses a traditional Chinese medicine preparation for freshwater fish baths, overcoming the shortcomings of conventional fish bath preparations that lose nutrients upon entering the water. It allows the medicine to form microcapsules and effectively adhere to the fish's surface, thus prolonging the residence and release time of the medicine on the fish's surface. This invention creatively encapsulates drugs for treating bacterial diseases and water mold in fish within microcapsules formed from extracts of Bletilla striata and elm bark. Simultaneously, the extracts of Bletilla striata and elm bark act as natural plant adhesives, forming a microfilm that effectively adheres to the fish's surface. This not only works during the bath but also adheres to the fish's surface after entering the water, forming a transparent, breathable, and elastic protective layer on the skin, acting as a physical barrier. The medicine is slowly released through the micropores on the microcapsules or by the slow degradation and rupture of the capsules. Surprisingly, the microcapsules adhere more strongly and more effectively to areas of scale loss and abrasions, while their adhesion to normal fish scales and gills is relatively weak. This characteristic makes them particularly suitable for the prevention and treatment of fish diseases caused by external injuries. The microcapsules, formed by the rational combination of extracts from Bletilla striata and elm bark, have good biocompatibility, biodegradability, and breathability with the fish skin. They are abundant, diverse, and renewable resources, non-irritating and non-toxic to the skin, and environmentally safe. They also feature functional sustained-release and local adhesive retention properties, allowing the drug to exert its therapeutic effect more effectively.

[0050] This invention discloses a traditional Chinese medicine preparation for freshwater fish baths, which effectively inhibits and kills various pathogens in freshwater fish, and has preventive and therapeutic effects on various fish diseases. The preparation consists of drug A and drug B. Drug A focuses on the sustained preventive and therapeutic effects on fish diseases after they enter large bodies of water, while drug B focuses on the rapid killing of pathogens on the fish surface and in the water. The combined use of both drugs results in a significant synergistic effect and better overall efficacy. Drug B contains rhubarb, sophora flavescens, and gallnut, all traditional fish medicines rich in flavonoids, alkaloids, sterols, glycosides, and other bioactive substances. In practice, these drugs have been widely used in the prevention and treatment of hemorrhagic septicemia and saprolegniasis caused by Aeromonas hydrophila, Aeromonas sobria, and Saprolegnia, with good results. The herbal preparation A for freshwater fish medicated baths contains abundant flavonoids, alkaloids, sterols, glycosides, and other bioactive substances, as well as rich secondary metabolites. It is a library of compounds that act on multiple targets, and can prevent and treat various aquatic fish diseases depending on the properties of different drugs and the desired effects.

[0051] This invention discloses a traditional Chinese medicine preparation for freshwater fish baths, which has astringent, hemostatic, and wound-healing effects, and is non-irritating to the skin. With proper combination, its effects are enhanced. The ingredients in this freshwater fish bath preparation, such as Bletilla striata, Galla chinensis, and Paeonia suffruticosa, have astringent, hemostatic, and wound-healing effects, rapidly restoring the fish's natural barrier function. Furthermore, after external application, it forms a film to protect the wound, reduce inflammation and irritation, and its astringent effect coagulates surface proteins, constricts local blood vessels, reduces congestion and exudation, and promotes wound healing without affecting normal skin physiological function, thus helping fish recover their health.

[0052] The freshwater fish medicated bath preparation described in this invention uses common Chinese herbal medicines available on the market. These herbs are green, natural, inexpensive, and readily available. When added to feed, they do not accumulate, leave no residue, or pollute the water. Attached Figure Description

[0053] Figure 1 The image shows a physical representation of the medicinal bath preparation made according to this invention.

[0054] Figure 2 This is a photograph of the protective layer formed when the microcapsules of the medicinal bath preparation described in this invention adhere to the surface of the fish during use. Detailed Implementation

[0055] The present invention will be further illustrated below with reference to specific embodiments. It should be understood that these embodiments are for illustrative purposes only and are not intended to limit the scope of the invention. Furthermore, it should be understood that after reading the teachings of this invention, those skilled in the art can make various alterations or modifications to the invention, and these equivalent forms also fall within the scope defined by the appended claims.

[0056] To avoid repetition, the sources of raw materials and processing techniques involved in the specific embodiments of the present invention will be explained in a unified manner as follows:

[0057] In the following embodiments, the Chinese herbal medicines used are all common varieties of Chinese herbal medicines on the market, and there are no special requirements.

[0058] In the following embodiments, the preparation method of the Bletilla striata extract is as follows: Bletilla striata tubers are dried in an oven at 70-80℃ for 2 hours, pulverized to pass through a 65-mesh sieve, placed in an extraction tank with a stirring device, water is added at a ratio of 1:15, and the mixture is extracted at 50-60℃ for 2 hours. The filtrate and residue are obtained by suction filtration. The filtrate is retained, and the residue (wet weight) is extracted three times in the same way. The three filtrates are combined, filtered to remove the residue, and the filtrate is concentrated to 1 / 3 of the original volume. Anhydrous ethanol is added at a volume ratio (anhydrous ethanol: extract) of 3:1, and the mixture is precipitated for 3 hours. The precipitate is separated by centrifugation at 6000 r / min for 15 minutes. The precipitate is dried, pulverized, and the Bletilla striata extract is obtained.

[0059] In the following embodiments, the preparation method of the elm bark extract is as follows: take elm bark, remove the outer bark, dry it in an oven at 70-80℃ for 2 hours, pulverize it through a 65-mesh sieve, put it into an extraction tank with a stirring device, add water at a ratio of 1:10, extract at 50-60℃ for 2 hours, filter to obtain filtrate and residue, retain the filtrate, and extract the residue (wet weight) three times in the same way, combine the three filtrates, filter to remove the residue, concentrate the filtrate to 1 / 3 of the original volume, introduce the filtrate into an equal volume of 95% ethanol, precipitate with alcohol for 3 hours, centrifuge at 6000 r / min for 15 minutes, separate the precipitate, dry the precipitate, pulverize it, and obtain the elm bark extract;

[0060] In the following embodiments, the preparation methods of the Isatis indigotica leaf extract, Salvia miltiorrhiza extract, and Paeonia suffruticosa root bark extract are as follows: Impurities in the Isatis indigotica leaves are removed, dried, pulverized through a 65-mesh sieve, and mixed evenly to obtain Isatis indigotica leaf powder; the Isatis indigotica leaf powder and anhydrous ethanol solution are mixed at a weight-to-volume ratio of 1:10, and extracted for 50-70 minutes under the conditions of ultrasonic power 250-350W, 2.5-3.5W / ml, frequency 20-40kHz, temperature 45-55℃, and stirring speed 80-120rpm / min; the mixture is then filtered to obtain filtrate and residue; the residue is extracted twice more under the same conditions; the residue is discarded, and the filtrates obtained from the three extractions are combined; the Isatis indigotica leaf filtrate is concentrated, freeze-dried, and pulverized to obtain the Isatis indigotica leaf extract; the preparation methods of the Paeonia suffruticosa root bark extract and Salvia miltiorrhiza extract are the same as those for the Isatis indigotica leaf extract.

[0061] In the following embodiments, the preparation methods of the Scutellaria baicalensis extract, Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract, and Gleditsia sinensis thorn extract are as follows: Remove impurities from Scutellaria baicalensis, dry it in the sun, and pulverize it through a 65-mesh sieve to obtain Scutellaria baicalensis powder; mix the Scutellaria baicalensis powder and 50% ethanol solution at a weight-to-volume ratio of 1:10, and extract for 50-70 minutes under the conditions of ultrasonic power 250-350W, 2.5-3.5W / ml, frequency 20-40kHz, temperature 45-55℃, and stirring speed 80-120rpm / min; filter to obtain filtrate and residue; extract the residue twice more under the same conditions; discard the residue and combine the filtrates obtained from the three extractions; concentrate the filtrate, freeze-dry it, and pulverize it to obtain Scutellaria baicalensis extract; the preparation methods of Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract, and Gleditsia sinensis thorn extract are the same as those for Scutellaria baicalensis extract.

[0062] In the following embodiments, the preparation method of the Pseudolarix amabilis bark extract is as follows: Impurities in the Pseudolarix amabilis bark are removed, the bark is dried, and pulverized through a 65-mesh sieve to obtain Pseudolarix amabilis bark powder; the Pseudolarix amabilis bark powder and 75% ethanol solution are mixed at a weight-to-volume ratio of 1:10, and extracted for 50-70 minutes under the conditions of ultrasonic power 250-350W, 2.5-3.5W / ml, frequency 20-40kHz, temperature 45-55℃, and stirring speed 80-120rpm / min; the mixture is then filtered to obtain the filtrate and residue; the residue is extracted twice more under the same conditions; the residue is discarded, and the filtrates obtained from the three extractions are combined; the filtrate is concentrated, freeze-dried, and pulverized to obtain the Pseudolarix amabilis bark extract.

[0063] In the following embodiments, the method of using the freshwater fish bath herbal preparation is to dissolve the freshwater fish bath herbal preparation in water at a concentration of 120-180 mg / L and bathe the fish for 20-40 minutes.

[0064] The freshwater fish herbal bath preparation provided by this invention utilizes microcapsule technology, overcoming the shortcomings of current freshwater fish bath preparations that fail to provide continuous protection to fish after they enter the aquaculture water. This allows the drug to form microcapsules that effectively adhere to the fish's surface, creating a transparent, breathable, and elastic protective layer on the skin, acting as a physical barrier. This prolongs the drug's residence time on the fish's surface and provides a slow-release effect. It also has astringent, hemostatic, and wound-healing properties, inhibits and kills various pathogens in freshwater fish, and has preventative and therapeutic effects on various fish diseases. It is green, safe, and environmentally friendly, and is particularly suitable for the recovery and healing of wounds on the surface of freshwater fish, as well as for preventing bacterial and fungal infections caused by injury.

[0065] Example 1

[0066] Step 1: Mix Bletilla striata extract and Ulmus pumila bark extract in a weight ratio of 20:1, add water, stir, and dissolve completely to prepare a 5% solution, which is denoted as Solution I.

[0067] Step 2: Take 200 parts by weight of Isatis indigotica leaf extract, 200 parts by weight of Salvia miltiorrhiza extract, 70 parts by weight of Paeonia suffruticosa root bark extract, 200 parts by weight of Scutellaria baicalensis extract, 100 parts by weight of Phellodendron chinense bark extract, 70 parts by weight of Coptis chinensis extract, 100 parts by weight of Angelica dahurica extract, 80 parts by weight of Gleditsia sinensis thorn extract, and 120 parts by weight of Pseudolarix amabilis bark extract, mix them into extract powder, and dissolve the obtained extract powder in an 80% ethanol solution at a weight-volume ratio (g:ml) of 1:2. Homogenize under high pressure at 20MPa for 10 minutes, and record the resulting solution as solution II.

[0068] Step 3: Take 2500 parts of solution I obtained in step 1 by volume, heat to 60°C, slowly add 100 parts of solution II obtained in step 2 while stirring. After the solution is completely added, continue stirring at a constant temperature of 60°C for 40 minutes.

[0069] Step 4: Homogenize the solution obtained in Step 3 under high pressure at a pressure of 30 MPa for 30 min to obtain solution III.

[0070] Step 5: Spray dry solution III at an inlet temperature of 160°C and an outlet temperature of 80°C to obtain microcapsule drug A.

[0071] Step 6: Dry rhubarb, sophora flavescens, and gallnut in an oven at 70℃ for 2 hours, then pulverize them separately to pass through a 65-mesh sieve. Take 300 parts of rhubarb, 150 parts of sophora flavescens, and 150 parts of gallnut by weight, and put them into an extraction tank equipped with a stirring device. Add water at a weight-to-volume ratio (g:ml) of 1:15, heat, and control the temperature at 70℃ for 2 hours. Filter to obtain filtrate and residue. Retain the filtrate, and extract the residue (wet weight) three times in the same way. Combine the three filtrates, filter to remove the residue, concentrate the filtrate, dry it, and pulverize it to pass through a 65-mesh sieve to obtain drug B.

[0072] Step 7: Mix medicine A and medicine B in a weight ratio of 1:3 to obtain the traditional Chinese medicine preparation for freshwater fish bath.

[0073] Example 2

[0074] Step 1: Mix Bletilla striata extract and Ulmus pumila bark extract in a weight ratio of 20-30:1, add water, stir, and dissolve completely to prepare a 7% solution, which is denoted as Solution I.

[0075] Step 2: Take 250 parts by weight of Isatis indigotica leaf extract, 175 parts by weight of Salvia miltiorrhiza extract, 85 parts by weight of Paeonia suffruticosa root bark extract, 175 parts by weight of Scutellaria baicalensis extract, 85 parts by weight of Phellodendron chinense bark extract, 85 parts by weight of Coptis chinensis extract, 85 parts by weight of Angelica dahurica extract, 100 parts by weight of Gleditsia sinensis thorn extract, and 100 parts by weight of Pseudolarix amabilis bark extract, mix them into extract powder, and dissolve the obtained extract powder in a 75% ethanol solution at a weight-volume ratio (g:ml) of 2:5. Homogenize under high pressure at 25MPa for 7 minutes, and record the resulting solution as solution II.

[0076] Step 3: Take 3000 parts of solution I obtained in step 1 by volume, heat to 55°C, slowly add 100 parts of solution II obtained in step 2 while stirring, and continue stirring at 55°C for 30 minutes after the solution has been completely added.

[0077] Step 4: Homogenize the solution obtained in Step 3 under high pressure at a pressure of 25 MPa for 35 min to obtain solution III.

[0078] Step 5: Spray dry solution III at an inlet temperature of 170°C and an outlet temperature of 90°C to obtain microcapsule drug A.

[0079] Step 6: Dry rhubarb, sophora flavescens, and gallnut in an oven at 75℃ for 2 hours, then pulverize them separately to pass through a 65-mesh sieve. Take 400 parts of rhubarb, 125 parts of sophora flavescens, and 125 parts of gallnut by weight, and put them into an extraction tank equipped with a stirring device. Add water at a weight-to-volume ratio (g:ml) of 1:15, heat, and control the temperature at 75℃ for 2 hours. Filter to obtain filtrate and residue. Retain the filtrate, and extract the residue (wet weight) three times in the same way. Combine the three filtrates, filter to remove the residue, concentrate the filtrate, dry it, and pulverize it to pass through a 65-mesh sieve to obtain drug B.

[0080] Step 7: Mix medicine A and medicine B in a weight ratio of 1:4 to obtain the traditional Chinese medicine preparation for freshwater fish bath.

[0081] Example 3

[0082] Step 1: Mix Bletilla striata extract and Ulmus pumila bark extract in a weight ratio of 30:1, add water, stir, and dissolve completely to prepare a 10% solution, which is denoted as Solution I.

[0083] Step 2: Take 300 parts by weight of Isatis indigotica leaf extract, 150 parts by weight of Salvia miltiorrhiza extract, 100 parts by weight of Paeonia suffruticosa root bark extract, 150 parts by weight of Scutellaria baicalensis extract, 70 parts by weight of Phellodendron chinense bark extract, 100 parts by weight of Coptis chinensis extract, 70 parts by weight of Angelica dahurica extract, 120 parts by weight of Gleditsia sinensis thorn extract, and 80 parts by weight of Pseudolarix amabilis bark extract, mix them into extract powder, and dissolve the obtained extract powder in a 70% ethanol solution at a weight-volume ratio (g:ml) of 1:3. Homogenize under high pressure at 30MPa for 5 minutes, and record the resulting solution as solution II.

[0084] Step 3: Take 3500 parts of the solution obtained in Step 1 by volume, heat to 50°C, slowly add 100 parts of the solution obtained in Step 2 while stirring. After the solution is completely added, continue stirring at a constant temperature of 50°C for 20 minutes.

[0085] Step 4: Homogenize the solution obtained in Step 3 under high pressure at a pressure of 20 MPa for 40 min to obtain solution III.

[0086] Step 5: Spray dry solution III at an inlet temperature of 180°C and an outlet temperature of 95°C to obtain microcapsule drug A.

[0087] Step 6: Dry rhubarb, sophora flavescens, and gallnut in an oven at 80℃ for 2 hours, then pulverize them separately to pass through a 65-mesh sieve. Take 500 parts of rhubarb, 150 parts of sophora flavescens, and 150 parts of gallnut by weight and place them in an extraction tank equipped with a stirring device. Add water at a weight-to-volume ratio (g:ml) of 1:15, heat, and control the temperature at 80℃ for 2 hours. Filter to obtain filtrate and residue. Retain the filtrate and extract the residue (wet weight) three times in the same way. Combine the three filtrates, filter to remove the residue, concentrate the filtrate, dry it, and pulverize it to pass through a 65-mesh sieve to obtain drug B.

[0088] Step 7: Mix medicine A and medicine B in a weight ratio of 1:5 to obtain the traditional Chinese medicine preparation for freshwater fish bath.

[0089] Comparative Example 1

[0090] The steps of this comparative example are basically the same as those of Example 2, except that:

[0091] Steps 6 and 7 are missing; the resulting product is microcapsule drug A, but drug B is missing.

[0092] Comparative Example 2

[0093] The steps of this comparative example are basically the same as those of Example 2, except that:

[0094] Steps 1-5 and step 7 are missing, resulting in drug B, but microcapsule drug A is missing.

[0095] Comparative Example 3

[0096] The steps of this comparative example are basically the same as those of Example 2, except that:

[0097] Steps 4 and 5 are missing, and drug A has not been microencapsulated.

[0098] Comparative Example 4

[0099] The steps of this comparative example are basically the same as those of Example 2, except that:

[0100] Step 1: Add the Bletilla striata extract to water according to the weight proportions, stir, and dissolve fully to prepare a 7% solution; elm bark extract is not available.

[0101] Example of effect

[0102] <Safety Experiment of Traditional Chinese Medicine Preparations for Freshwater Fish Baths>

[0103] (1) Basic feed for Pengze crucian carp: fish meal 15.00%, soybean meal 25.00%, rapeseed meal 15.00%, cottonseed meal 15.00%, wheat middlings 20.00%, rice bran 3.00%, corn gluten meal 4.1%, vitamin and mineral premix 1%, salt 0.4%, calcium dihydrogen phosphate 1.5%.

[0104] (2) Experimental fish: Select Pengze crucian carp with a size of (52±5) grams, put them in a glass aquarium with temperature control and oxygenation equipment, and temporarily keep them for 7 days. During the temporary keeping period, feed them basic feed.

[0105] (3) Safety test

[0106] After the temporary rearing period, once the Pengze crucian carp had adapted to the aquarium environment, they were immediately divided into 5 groups. Control group: no medicated bath; Low-dose group: medicated bath with 120 mg / L of the herbal preparation (minimum effective dose); High-dose group: medicated bath with 180 mg / L of the herbal preparation (maximum effective dose); 2x group: medicated bath with 360 mg / L of the herbal preparation (twice the maximum effective dose); 4x group: medicated bath with 720 mg / L of the herbal preparation (four times the maximum effective dose). All medicated bath groups received one bath per day for 30 minutes each time for 5 consecutive days. During the experiment, they were fed normally twice a day, until they were full with a slight surplus.

[0107] Table 1. Safety of Traditional Chinese Medicine Preparations for Freshwater Fish Baths on Pengze Crucian Carp

[0108]

[0109] After the medicated bath, all experimental groups survived without any deaths. Dissection of the Pengze crucian carp in each experimental group revealed no significant differences in the appearance, color, elasticity, or intestinal wall thickness of the liver, spleen, intestines, and other internal organs. Even when the dosage of the medicated bath preparation exceeded four times the maximum effective dose (4-fold group), there were no significant changes in the internal organs, indicating that the obtained medicated bath preparation is safe and reliable when used within the effective dosage range, with no toxic side effects.

[0110] <The therapeutic effects of traditional Chinese medicine preparations in freshwater fish baths on external injuries>

[0111] (1) Experimental fish: Same as safety experiment.

[0112] (2) Wound modeling: After the temporary rearing period, when the Pengze crucian carp adapts to the aquarium environment, external injuries such as scale loss and abrasions are created using fishing nets. The Pengze crucian carp have varying degrees of scale loosening and falling off, as well as congestion on the upper and lower jaws, eyeballs, gill covers, caudal peduncle, fin base and abdomen.

[0113] (3) The therapeutic effect of traditional Chinese medicine preparations for freshwater fish baths on external injuries of Pengze crucian carp

[0114] The Pengze crucian carp were randomly divided into 9 groups, with 2 replicates per group. Control group: no trauma, no medicated bath; Positive control group: trauma, no medicated bath; Medicated bath drug A group: trauma, using only drug A (45 mg / L) from the medicated bath preparation, without drug B, prepared as Comparative Example 1; Medicated bath drug B group: trauma, using only drug B (150 mg / L) from the medicated bath preparation, without drug A, prepared as Comparative Example 2; Medicated bath without microencapsulation group: trauma, using the non-microencapsulated medicated bath preparation (150 mg / L), prepared as Comparative Example 3; Medicated bath without elm bark group: trauma, using the medicated bath preparation without elm bark (150 mg / L), prepared as Comparative Example 4. Low-dose medicated bath group: trauma, medicated bath with Chinese herbal preparation 120mg / L, the lowest effective dose; Medium-dose medicated bath group: trauma, medicated bath with Chinese herbal preparation 150mg / L, the middle effective dose; High-dose medicated bath group: trauma, medicated bath with Chinese herbal preparation 180mg / L, the highest effective dose; All medicated bath groups, medicated bath for 30 minutes, medicated bath once, normal feeding during the experiment, feeding twice a day until full with a slight surplus.

[0115] Table 2. The therapeutic effects of traditional Chinese medicine preparations used in freshwater fish baths on external injuries of Pengze crucian carp.

[0116]

[0117] Note: Different shoulder note letters in the same column indicate significant differences (P<0.05), and if any shoulder note number is the same, the difference is not significant (P>0.05).

[0118] All members of the control group without wound modeling survived. The survival rate of the positive control group without medicated bath treatment after wound modeling was 52.5%, indicating successful model establishment. After wound modeling, most Pengze crucian carp exhibited stress responses, manifesting as slow movement, restlessness, and susceptibility to fright. They also showed varying degrees of scale loosening and loss, as well as congestion on the upper and lower jaws, eyes, gill covers, caudal peduncle, fin bases, and abdomen. Compared to the control group, the Pengze crucian carp that survived the medicated bath showed increased vitality, faster swimming, and normal body color. They also resumed grouping and were less easily frightened. In some individuals, the congestion symptoms on the upper and lower jaws, gill covers, eyes, caudal peduncle, and fin bases disappeared, and the scales became compact again. Areas where scales had fallen off were smoother, darker in color, and closer to the body color. In more severely injured individuals, the congestion symptoms were significantly alleviated, and the color of the wound changed from dark red to light red and then to brown. The wounds where scales had fallen off began to heal.

[0119] Except for the group treated with medicated bath drug B, which showed no significant difference from the positive control group (P>0.05), all other experimental groups were significantly higher than the positive control group (P<0.05), indicating that the use of drug A alone, or the combination of drug A and drug B, could significantly improve the survival rate (P<0.05). Drug B can inhibit or kill pathogens on the surface of fish during medicated baths. However, after the medicated bath, when the Pengze crucian carp enter normal water bodies, it cannot provide effective protection for the fish. Aeromonas hydrophila, Aeromonas sobria, and Saprolegnia spores are resident bacteria in normal water bodies. When the fish's protective barrier is intact and the concentration of pathogens is low, serious diseases generally do not occur. However, when scales are lost, injuries occur, or the concentration of pathogens is too high due to netting, transportation, etc., or when the fish's immunity is reduced, various diseases may break out, and even large-scale deaths may occur. The survival rate of the medium and high doses of the medicinal herbs in the bath was significantly higher than that of the medicinal herb group B and the non-microencapsulated medicinal herb group (P<0.05), indicating that the survival rate was significantly improved after microencapsulation treatment.

[0120] <Preventive and therapeutic effects of traditional Chinese medicine preparations for freshwater fish baths on hemorrhagic septicemia in crucian carp>

[0121] (1) Experimental fish: Same as safety experiment.

[0122] (2) Preparation of infecting bacterial strains

[0123] Aeromonas hydrophila was inoculated into beef extract peptone broth and incubated at 28°C for 24 hours. The bacterial concentration was determined by turbidimetric assay, and the viable count was adjusted to 8 × 10⁶ using sodium chloride injection. 7 per mL.

[0124] (3) Wound modeling: Same as Pengze crucian carp trauma experiment.

[0125] (4) Prevention and treatment of hemorrhagic septicemia in crucian carp in Pengze

[0126] In the prevention experiment, Pengze crucian carp were bathed in a medicated solution for 30 minutes, then placed in a glass aquarium equipped with temperature control and aeration equipment. 20 ml of Aeromonas hydrophila solution was added per cubic meter of water. In the treatment experiment, Pengze crucian carp were temporarily kept in water with 20 ml of Aeromonas hydrophila solution added per cubic meter of water for 1-3 days. At least one-fifth of the fish showed typical symptoms of hemorrhagic septicemia before receiving a medicated bath. The prevention experiment involved one medicated bath, while the treatment experiment involved one medicated bath daily for three consecutive days, each lasting 30 minutes. During the experiments, the fish were fed normally twice daily, until they were slightly satiated with some food remaining.

[0127] Both the prevention and treatment trials of Pengze crucian carp were conducted using randomized grouping. The fish were randomly divided into 9 groups, each with a replicate, and each group contained 20 fish. The control group consisted of no trauma, no infection, and no medicated bath treatment. The positive control group consisted of trauma and infection, and no medicated bath treatment. The medicated bath drug A group consisted of trauma and infection, using only drug A (45 mg / L) from the medicated bath preparation, without drug B (as in Comparative Example 1). The medicated bath drug B group consisted of trauma and infection, using only drug B (150 mg / L) from the medicated bath preparation, without drug A (as in Comparative Example 2). The non-microencapsulated medicated bath group consisted of trauma and infection, using the non-microencapsulated medicated bath preparation (150 mg / L) (as in Comparative Example 3). The medicated bath lacking elm bark group consisted of trauma and infection, using the medicated bath preparation without elm bark (150 mg / L) (as in Comparative Example 4). Low-dose medicated bath group: trauma, infection, medicated bath Chinese medicine preparation 120mg / L, the lowest effective dose; Medium-dose medicated bath group: trauma, infection, medicated bath Chinese medicine preparation 150mg / L, the middle effective dose; High-dose medicated bath group: trauma, infection, medicated bath Chinese medicine preparation 180mg / L, the highest effective dose.

[0128] Table 3. Preventive effect of traditional Chinese medicine preparations for freshwater fish baths on hemorrhagic septicemia in crucian carp from Pengze.

[0129]

[0130]

[0131] Note: Different shoulder note letters in the same column indicate significant differences (P<0.05), and if any shoulder note number is the same, the difference is not significant (P>0.05).

[0132] Table 4. Therapeutic effects of traditional Chinese medicine preparations used in freshwater fish baths on hemorrhagic septicemia in crucian carp from Pengze.

[0133]

[0134] Note: Different shoulder note letters in the same column indicate significant differences (P<0.05), and if any shoulder note number is the same, the difference is not significant (P>0.05).

[0135] No deaths occurred in the control group during the experiment. In the prevention test, a wound was artificially created in the positive control group and 20 ml / m² of fluid was introduced. 3The survival rate of Aeromonas hydrophila in the water was 22.5%, indicating that the model was successfully established. In the positive control group, most Pengze crucian carp showed symptoms within 1-3 days. Mild congestion was initially observed in the mouth, upper and lower jaws, gill covers, and base of the pectoral fins of the diseased fish, with punctate congestion on the abdomen and sides of the body. As the disease progressed, congestion and congestion around the eyes and eye sockets occurred, the eyeballs bulged, bleeding on the body surface intensified, the abdomen swelled, and the anus became red and swollen. Dissection revealed congestion and hemorrhage in multiple internal organs such as the gills, liver, kidneys, and intestines, which subsequently became lighter in color due to blood loss. The liver was yellowish-brown, and the spleen and kidneys were significantly enlarged, lost their original elasticity, and were easily broken. The intestinal wall was congested, and the abdominal cavity contained a large amount of bloody ascites, which was dark red in color and had low viscosity. The vast majority of deaths occurred within 5 days, indicating that Pengze crucian carp hemorrhagic septicemia is a rapidly developing and explosive disease.

[0136] The survival rates of crucian carp in the low, medium, and high dose groups were significantly higher than those in the positive control group (P<0.05), with no significant differences among the three groups (P>0.05). Higher dosages resulted in better preventative effects, exhibiting a dose-response effect. Specifically, the survival rates of the low-dose, medium-dose, and high-dose medicated bath groups were 52.5%, 57.5%, and 65% higher than the positive control group, respectively. The low, medium, and high dose groups represented the minimum, medium, and maximum doses of the medicated bath medicine, indicating that standard dosage has a significant preventative effect against hemorrhagic septicemia in crucian carp in Pengze. The survival rates of the medicated bath medicine A group, medicine B group, and the non-microencapsulated medicated bath group were significantly lower than those of the medium-dose and high-dose groups (P<0.05), indicating that the absence of medicine A or B, or the lack of microencapsulation, significantly affected the efficacy of the medicated bath medicine. The survival rate of the medicated bath group lacking elm bark was lower than that of the low, medium, and high dose groups, but the difference was not statistically significant (P>0.05), indicating that the lack of elm bark extract in drug A also affected the efficacy of the medicated bath preparation to some extent. The results of the treatment and prevention experiments were similar. Compared with the positive control group, the survival rates of the low-dose, medium-dose, and high-dose medicated bath groups were increased by 42.5%, 45%, and 50%, respectively. The survival rates of all three medicated bath groups were lower than those of the corresponding prevention groups, indicating that the medicated bath treatment was more effective in preventing hemorrhagic septicemia in Pengze crucian carp than in treating it.

[0137] The rhubarb, sophora flavescens, and gallnut in the herbal preparation B for freshwater fish medicated baths are all traditional fish medicines, rich in flavonoids, alkaloids, sterols, glycosides, and other bioactive substances. In practice, they have been widely used in the prevention and treatment of hemorrhagic septicemia and saprolegniasis caused by Aeromonas hydrophila, Aeromonas sobria, and Saprolegnia, with good results. However, a drawback is that after the fish are placed back into normal water, the medication on their body surface dissipates quickly. Aeromonas hydrophila, Aeromonas sobria, and Saprolegnia spores are resident bacteria in normal water. When the fish's protective barrier is intact and the concentration of pathogens is low, they generally do not cause serious diseases. However, when scales are lost due to netting or transportation, injuries occur, or the concentration of pathogens is too high, or the fish's immunity is reduced, various diseases can break out, even leading to large-scale mortality. The herbal preparation A for freshwater fish medicated baths contains abundant flavonoids, alkaloids, sterols, glycosides, and other bioactive substances, including Isatis tinctoria leaf, Salvia miltiorrhiza, Paeonia suffruticosa root bark, Scutellaria baicalensis root, Phellodendron chinense bark, Coptis chinensis root, Angelica dahurica root, Gleditsia sinensis thorns, and Pseudolarix amabilis bark. It is rich in secondary metabolites and represents a library of compounds acting on multiple targets. Furthermore, based on the different properties of various drugs and the desired effects, multiple combinations offer endless possibilities for the efficacy and variety of traditional Chinese medicine. Different combinations can produce unexpected effects, preventing and treating various aquatic fish diseases. Bletilla striata and elm bark possess astringent, hemostatic, anti-inflammatory, analgesic, and wound-healing properties, and are also excellent biological adhesives. Through homogenization and spray drying processes, the active ingredients of extracts from Isatis indigotica, Salvia miltiorrhiza, Paeonia suffruticosa, Scutellaria baicalensis, and Phellodendron chinense are encapsulated in microcapsules composed of Bletilla striata and elm bark, adhering to the surface of fish. This forms a transparent, breathable, and elastic protective layer on the skin, acting as a physical barrier. This layer can remain in the normal water environment for a certain period, promoting wound healing and continuously exerting its medicinal effects, overcoming the shortcomings of conventional medicated bath preparations, such as short-lived efficacy and loss upon entering the water. Furthermore, the adhesive components of Bletilla striata and elm bark are mainly composed of polysaccharides and glycoproteins, which are biodegradable. Experiments have shown that they are particularly effective at adhering to the epidermis in areas with scale loss and abrasions, possibly due to reduced mucus in these areas. They exhibit good biocompatibility and will not affect the normal physiological functions of fish within the medicinal dosage range.

[0138] <Preventive Effect of Traditional Chinese Medicine Preparations in Freshwater Fish Baths on Saprolegniasis in Carp Eggs>

[0139] (1) Fish eggs

[0140] The fish eggs were laid by the same batch of carp (2 males and 2 females), with an average fertilization rate of 83%.

[0141] (2) The preventive effect of traditional Chinese medicine preparations in medicated baths on saprolegniasis in carp eggs

[0142] Fish eggs were divided into three groups, with three replicates per group and 2000 eggs per replicate. The specific groups were: negative control group (4 mg / L malachite green bath); positive control group (no bath); and the group receiving a traditional Chinese medicine bath (150 mg / L traditional Chinese medicine). After egg formation, the fish eggs were bathed once for 30 minutes. Following the bath, the eggs were placed in an incubation tank using well water, which was fully aerated in a storage tank. All indicators were calculated after the experiment.

[0143] (3) Indicator Calculation

[0144] Hatching rate (%) = Number of hatched fry / Total number of hatched eggs × 100

[0145] Saprolegnia mortality rate (%) = Number of eggs killed by Saprolegnia / Total number of hatched fish eggs × 100

[0146] Mortality rate due to other factors (%) = Number of eggs killed by other factors / Total number of hatched eggs × 100

[0147] Deformity rate (%) = Number of deformed fry / Number of hatched fry × 100

[0148] Table 5. Preventive effect of traditional Chinese medicine preparations used in freshwater fish baths on saprolegniasis in carp eggs.

[0149]

[0150] Note: Different shoulder note letters in the same column indicate significant differences (P<0.05); if any shoulder note number is the same, the difference is not significant (P>0.05). Data in the table are (mean ± standard error).

[0151] The mortality rate of Saprolegnia in the malachite green group and the medicated bath group was significantly lower than that in the positive control group without medicated bath treatment (P<0.05), while there was no significant difference between the two groups (P>0.05). Specifically, the mortality rate of Saprolegnia in the medicated bath group was reduced by 27.28% compared to the positive control group. This indicates that Saprolegnia is a serious disease during fish egg hatching, and that soaking fish eggs in malachite green and medicated bath treatments significantly reduces the incidence of Saprolegnia. Notably, malachite green treatment significantly increased the deformity rate of fish eggs, indicating that malachite green, as a chemical agent, has a certain toxic effect on fish eggs. However, there was no significant difference in the deformity rate between the medicated bath group and the positive control group, suggesting that the overall effect of using medicated bath treatments to prevent Saprolegnia in fish eggs is superior to that of malachite green.

[0152] <Pond Aquaculture Experiment>

[0153] Between April and July 2023, a large fishing pond in Luoyang purchased various aquatic fish from other places and transported them to the pond for fishing. Some of the fish were disinfected with conventional salt and potassium permanganate, while others were disinfected with the medicated bath preparation of traditional Chinese medicine prepared in Example 2 of this invention at a concentration of 150 mg / L. All fish were disinfected once and released into the pond after 20-40 minutes of disinfection. The results were as follows.

[0154] Table 6. Practical Application Effects of Traditional Chinese Medicine Preparations for Freshwater Fish Baths

[0155]

[0156] Compared with conventional disinfectants such as salt and potassium permanganate, the mortality rate after bathing with the traditional Chinese medicine preparation of this invention is significantly reduced.

[0157] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A method for preparing a traditional Chinese medicine preparation for freshwater fish baths, characterized in that, Includes the following steps: S1. Mix Bletilla striata extract and Ulmus pumila bark extract at a weight ratio of 20-30:1, add water, stir, and obtain solution I with a concentration of 5-10% for later use; S2, according to the following weight proportions, take 200-300 parts of Isatis indigotica leaf extract, 150-200 parts of Salvia miltiorrhiza extract, 70-100 parts of Paeonia suffruticosa root bark extract, 150-200 parts of Scutellaria baicalensis extract, 70-100 parts of Phellodendron chinense bark extract, 70-100 parts of Coptis chinensis extract, 70-100 parts of Angelica dahurica extract, 80-120 parts of Gleditsia sinensis thorn extract and 80-120 parts of Pseudolarix amabilis bark extract, mix them into extract powder, dissolve the obtained extract powder in a 70-80% ethanol solution at a weight-volume ratio of 1:2-3, homogenize under high pressure to obtain solution II; S3. Take 2500-3500 parts of solution I according to volume fractions, heat to 50-60℃, add 100 parts of solution II while stirring, continue stirring at 50-60℃ for 20-40 minutes, homogenize under high pressure to obtain solution III, and spray dry solution III to obtain microcapsule drug A. S4. Take 300-500 parts of rhubarb powder, 100-150 parts of sophora flavescens powder and 100-150 parts of gallnut powder according to the weight ratio, extract, filter to remove the residue, concentrate the filtrate, dry and pulverize to obtain drug B. S5. Mix medicine A and medicine B in a weight ratio of 1:3-5 to obtain a traditional Chinese medicine preparation for freshwater fish bath. Specifically, step S4 involves drying rhubarb, sophora flavescens, and gallnut in an oven at 70-80℃, pulverizing them, taking 300-500 parts of rhubarb powder, 100-150 parts of sophora flavescens powder, and 100-150 parts of gallnut powder by weight, placing them in an extraction tank equipped with a stirring device, adding water at a weight-to-volume ratio of 1:15, heating, controlling the temperature at 70-80℃, extracting, filtering, concentrating the filtrate, drying, and pulverizing to obtain drug B, which is a mixture of rhubarb extract, sophora flavescens extract, and gallnut extract. The preparation method of the Bletilla striata extract is as follows: the Bletilla striata tuber is dried in an oven at 70-80℃, pulverized, extracted with water, filtered and concentrated to obtain an extract, then anhydrous ethanol is added to the extract, alcohol precipitation is performed, centrifugation is performed to separate the precipitate, drying is performed and pulverized to obtain the Bletilla striata extract. The preparation method of the elm bark extract is as follows: take elm bark, dry it in an oven at 70-80℃, pulverize it, add water to extract it, filter it, concentrate the filtrate to obtain the extract, introduce the extract into 95% ethanol, precipitate it with alcohol, centrifuge it, separate the precipitate, dry it, pulverize it to obtain the elm bark extract. The preparation methods of the Isatis tinctoria leaf extract, Salvia miltiorrhiza extract, and Paeonia suffruticosa root bark extract are as follows: Isatis tinctoria leaf powder, Salvia miltiorrhiza powder, Paeonia suffruticosa root bark powder and anhydrous ethanol solution are mixed separately, ultrasonically extracted, filtered, the filtrate is concentrated, freeze-dried, and pulverized to obtain Isatis tinctoria leaf extract, Paeonia suffruticosa root bark extract and Salvia miltiorrhiza extract. The preparation methods of the Scutellaria baicalensis extract, Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract, and Gleditsia sinensis thorn extract are as follows: Scutellaria baicalensis powder, Phellodendron chinense powder, Coptis chinensis powder, Angelica dahurica powder, Gleditsia sinensis thorn powder and 50% ethanol solution are mixed, ultrasonically extracted, filtered, the filtrate is concentrated, freeze-dried, and pulverized to obtain Scutellaria baicalensis extract, Phellodendron chinense extract, Coptis chinensis extract, Angelica dahurica extract and Gleditsia sinensis thorn extract; The preparation method of the Pseudolarix amabilis bark extract is as follows: mix Pseudolarix amabilis bark powder and 75% ethanol solution, extract by ultrasonication, filter, concentrate the filtrate, freeze dry, and pulverize to obtain Pseudolarix amabilis bark extract.

2. The method for preparing a freshwater fish bath traditional Chinese medicine preparation according to claim 1, characterized in that, In step S2, the pressure of high-pressure homogenization is 20MPa-30MPa, and the homogenization time is 5-10min; in step S3, the pressure of high-pressure homogenization is 20MPa-30MPa, and the homogenization time is 30-40min.

3. The method for preparing a freshwater fish medicated bath preparation according to claim 1, characterized in that, In step S3, the inlet temperature of the spray dryer is 160-180℃, and the outlet temperature is 80-95℃.

4. A traditional Chinese medicine preparation for freshwater fish baths obtained by any of the preparation methods described in claims 1-3.

5. The application of the freshwater fish medicated bath preparation as described in claim 4 in the preparation of drugs for the prevention and treatment of freshwater fish injuries, hemorrhagic septicemia, and saprolegniasis.

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