Traditional Chinese medicine composition for preventing diseases of aquatic animals, preparation method, breeding method and application

Abstract

The invention discloses a traditional Chinese medicine composition for preventing diseases of aquatic animals, a preparation method, a breeding method and application, and belongs to the technical field of aquaculture. The traditional Chinese medicine composition is prepared from the following raw materials in parts by weight: 10 to 20 parts of radix astragali seu hedysari, 5 to 15 parts of rhizoma dioscoreae, 5 to 15 parts of poria cocos, 10 to 30 parts of medicated leaven, 5 to 15 parts of fructus hordei germinatus, 10 to 20 parts of fruit peel, 10 to 20 parts of bran and 2 to 8 parts of liquorice root. The traditional Chinese medicine composition disclosed by the invention has the effects of invigorating spleen and replenishing qi, supporting healthy energy, promoting digestion and removing food stagnation, and resolving dampness and resisting stress, and can prevent diseases and improve the growth performance of the aquatic animals by enhancing the autoimmunity of the aquatic animals. Through the two-stage fermentation process, the synergistic effect of substrate degradation and functional flora can be fully exerted, the growth performance and disease resistance are remarkably improved, and the death rate after toxic attack is reduced.

Description

Technical Field

[0001] This invention belongs to the field of aquaculture technology, and particularly relates to a traditional Chinese medicine composition for preventing diseases in aquatic animals, its preparation method, its breeding method, and its application. Background Technology

[0002] As aquaculture develops rapidly towards large-scale and intensive operations, the stocking density continues to increase, leading to increasingly prominent problems such as water environment deterioration, frequent disease outbreaks, and decreased digestive and absorptive capacity of aquatic animals. These issues not only restrict the growth performance of aquatic animals and reduce the survival rate of farmed animals, but also affect the quality of farmed products, resulting in serious economic losses.

[0003] Currently, the prevention and control of aquatic animal diseases mainly relies on chemical disinfectants and antibiotics. Long-term or improper use of chemical disinfectants can easily disrupt the microecological balance of aquaculture water and cause stress and toxic side effects in aquatic animals. Meanwhile, the overuse of antibiotics leads to a series of problems, including drug residues, antibiotic resistance in pathogens, and compromised quality and safety of farmed products. Finding safe, efficient, and sustainable disease prevention strategies has become an urgent need for the industry's development.

[0004] Traditional Chinese medicine (TCM) is considered a potential solution to replace or reduce antibiotic use due to its natural origin, multi-target effects, low risk of drug resistance, and low residue risk. Numerous studies have shown that many TCM herbs and their active ingredients possess multiple effects, including enhancing immunity, antibacterial and antiviral activity, antioxidant properties, and growth promotion. However, traditional methods of applying TCM herbs suffer from drawbacks such as the large molecular components in the raw materials being difficult for aquatic animals to digest and absorb efficiently, resulting in low bioavailability and insufficient dissolution of active ingredients, thus failing to fully realize their physiological regulatory effects and leading to less than expected practical application results.

[0005] Therefore, developing a traditional Chinese medicine preparation that can overcome the above limitations, enhance the resistance of aquatic animals, effectively prevent diseases, and promote growth is of great significance for promoting the green transformation and high-quality development of the aquaculture industry. Summary of the Invention

[0006] In view of this, the purpose of the present invention is to provide a traditional Chinese medicine composition for preventing diseases in aquatic animals, which can effectively improve the growth performance and disease resistance of aquatic animals.

[0007] Another object of the present invention is to provide a method for preparing the aforementioned traditional Chinese medicine composition.

[0008] Another object of the present invention is to provide the application of the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method in the preparation of products that promote the growth of aquatic animals and / or prevent aquatic animal diseases.

[0009] Another object of the present invention is to provide a method for preventing diseases in aquatic animals.

[0010] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a traditional Chinese medicine composition for preventing diseases in aquatic animals. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10-20 parts of Astragalus membranaceus, 5-15 parts of Dioscorea opposita, 5-15 parts of Poria cocos, 10-30 parts of Medicated Leaven, 5-15 parts of Malt, 10-20 parts of Pericarpium Citri Reticulatae, 10-20 parts of Wheat Bran, and 2-8 parts of Glycyrrhiza uralensis.

[0011] Preferably, the peel includes any one or more of the following: lychee peel, pineapple peel, banana peel, and dragon fruit peel.

[0012] The present invention also provides a method for preparing the traditional Chinese medicine composition, the method comprising: weighing each raw material according to the weight parts, crushing and sieving, mixing, and obtaining a mixture; inoculating the mixture with Bacillus subtilis bacterial solution, and sequentially carrying out a first stage of aerobic fermentation and a second stage of anaerobic fermentation; after the second stage of anaerobic fermentation is completed, drying to obtain the traditional Chinese medicine composition.

[0013] Preferably, the mesh size of the sieve is 40 to 80 mesh.

[0014] Preferably, before adding Bacillus subtilis bacterial solution to the mixture, the moisture content is further adjusted to 40%~50%.

[0015] Preferably, the viable cell concentration of the Bacillus subtilis bacterial solution is 1×10⁻⁶. 8 ~1×10 10 cfu / mL; the inoculation amount of the Bacillus subtilis bacterial solution is 0.3%~0.8% of the dry weight of the mixture.

[0016] Preferably, the conditions for aerobic fermentation include: fermentation temperature of 30~40℃ and time of 60~80h.

[0017] Preferably, during the aerobic fermentation process, the material is turned over once every 8 to 12 hours.

[0018] Preferably, the anaerobic fermentation conditions include: fermentation temperature of 25~35℃ and time of 96~144h.

[0019] The present invention also provides the application of the traditional Chinese medicine composition described herein or the traditional Chinese medicine composition prepared by the preparation method in the preparation of products that promote the growth of aquatic animals and / or prevent aquatic animal diseases.

[0020] The present invention also provides a method for preventing diseases in aquatic animals, the method comprising: mixing the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method with feed and feeding it to aquatic animals.

[0021] Preferably, the amount of the traditional Chinese medicine composition added is 0.5% to 3% of the feed mass.

[0022] The beneficial effects of this invention are: This invention's herbal composition uses Astragalus membranaceus as the principal ingredient, Dioscorea opposita, Poria cocos, Medicated Leaven, and Malt as assistant ingredients, Pericarpium Citri Reticulatae and Wheat Bran as adjuvant ingredients, and Glycyrrhiza uralensis as the guiding ingredient. The combined effects of these herbs are to invigorate the spleen and replenish qi, strengthen the body's resistance, promote digestion, eliminate dampness, and counteract adverse conditions. By enhancing the aquatic animals' own resistance, it prevents diseases and improves their growth performance. This invention utilizes a two-stage fermentation process to fully leverage the synergistic effect of substrate degradation and functional microbial communities, significantly improving growth performance and disease resistance, and reducing mortality after viral challenge. Applying this herbal composition to aquatic animal farming can effectively improve the growth performance and disease resistance of aquatic animals, and it has the advantages of being green and safe, leaving no harmful residues, and being less likely to induce drug resistance, providing a novel and efficient solution for disease prevention in aquatic animals. Detailed Implementation

[0023] This invention provides a traditional Chinese medicine composition for preventing diseases in aquatic animals. The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10-20 parts of Astragalus membranaceus, 5-15 parts of Dioscorea opposita, 5-15 parts of Poria cocos, 10-30 parts of Medicated Leaven, 5-15 parts of Malt, 10-20 parts of Pericarpium Citri Reticulatae, 10-20 parts of Wheat Bran, and 2-8 parts of Glycyrrhiza uralensis.

[0024] In this invention, the fruit peel preferably includes any one or more of the following: lychee peel, pineapple peel, banana peel, and dragon fruit peel.

[0025] In this invention, the weight percentage of Astragalus membranaceus is preferably 10-20 parts, more preferably 12-18 parts, and even more preferably 15 parts. The weight percentage of Dioscorea opposita is preferably 5-15 parts, more preferably 8-12 parts, and even more preferably 10 parts. The weight percentage of Poria cocos is preferably 5-15 parts, more preferably 8-12 parts, and even more preferably 10 parts. The weight percentage of Medicated Leaven is preferably 10-30 parts, more preferably 15-25 parts, and even more preferably 20 parts. The weight percentage of Malt is preferably 5-15 parts, more preferably 8-12 parts, and even more preferably 10 parts. The weight percentage of Pericarpium Citri Reticulatae is preferably 10-20 parts, more preferably 12-18 parts, and even more preferably 15 parts. The weight percentage of Wheat Bran is preferably 10-20 parts, more preferably 12-18 parts, and even more preferably 15 parts. The weight percentage of Glycyrrhiza uralensis is preferably 2-8 parts, more preferably 4-6 parts, and even more preferably 5 parts.

[0026] This invention uses Astragalus membranaceus as the principal ingredient, which invigorates the spleen and lungs, replenishes qi and strengthens the exterior, significantly enhancing the body's non-specific immunity and laying the foundation for resisting pathogen invasion. It uses Dioscorea opposita, Poria cocos, medicated leaven, and malt as assistant ingredients. Dioscorea opposita assists the principal ingredient in invigorating the spleen and stomach, replenishing qi and nourishing yin, and promoting nutrient absorption. Poria cocos invigorates the spleen, calms the mind, and promotes diuresis and dampness elimination, helping the principal ingredient invigorate the spleen and alleviate internal dampness and turbidity caused by water quality stress. Medicated leaven and malt promote digestion, resolve food stagnation, harmonize the stomach, and improve feed utilization. Active medicated leaven also provides a microbial basis for subsequent fermentation processes. It uses fruit peel and wheat bran as adjuvant ingredients. Fruit peel resolves turbidity and harmonizes the middle jiao, and also provides functional components such as polyphenols. Wheat bran harmonizes the middle jiao and benefits the stomach, serving as a fermentation carrier to loosen the medicinal substances and synergistically transform them. Both are adjuvant ingredients. Finally, it uses Glycyrrhiza uralensis as the guiding ingredient, harmonizing the various ingredients, detoxifying, harmonizing the middle jiao, and guiding the medicine to its proper channels. When used together, these herbs work synergistically to invigorate the spleen and replenish qi, strengthen the body's resistance, promote digestion and relieve stagnation, and eliminate dampness and counteract adverse conditions. By enhancing the aquatic animals' own resistance, they can prevent diseases and improve their growth performance.

[0027] The present invention also provides a method for preparing the traditional Chinese medicine composition, the method comprising: weighing each raw material according to the weight parts, crushing and sieving, mixing, and obtaining a mixture; inoculating the mixture with Bacillus subtilis bacterial solution, and sequentially carrying out a first stage of aerobic fermentation and a second stage of anaerobic fermentation; after the second stage of anaerobic fermentation is completed, drying to obtain the traditional Chinese medicine composition.

[0028] In this invention, the mesh size of the sieve is preferably 40-80 mesh, more preferably 50-70 mesh, and even more preferably 60 mesh.

[0029] In this invention, before adding Bacillus subtilis bacterial solution to the mixture, it is preferable to further adjust the moisture content; the moisture content is preferably adjusted to 40%~50%, more preferably 42%~47%, and even more preferably 45%.

[0030] In this invention, the viable cell concentration of the Bacillus subtilis bacterial solution is preferably 1×10⁻⁶. 8 ~1×10 10 cfu / mL, more preferably 5×10 8 ~5×10 9 cfu / mL, more preferably 1×10⁻⁶ 9 cfu / mL; the source of the Bacillus subtilis is not particularly limited and can be obtained through commercial channels; the preparation method of the Bacillus subtilis bacterial solution is not particularly limited, but in some embodiments, it preferably includes: dissolving the Bacillus subtilis bacterial agent in water to obtain a Bacillus subtilis bacterial solution with the corresponding live bacteria concentration.

[0031] In this invention, the inoculation amount of Bacillus subtilis bacterial solution is preferably 0.3% to 0.8% of the dry weight of the mixture, more preferably 0.4% to 0.6%, and even more preferably 0.5%.

[0032] In this invention, the temperature of the aerobic fermentation is preferably 30~40℃, more preferably 32~37℃, and even more preferably 35℃; the time of the aerobic fermentation is preferably 60~80h, more preferably 64~76h, and even more preferably 72h.

[0033] In this invention, during the aerobic fermentation process, the material is preferably turned over once every 8 to 12 hours, and more preferably once every 10 hours.

[0034] This invention utilizes the naturally abundant complex microbial systems of raw materials such as Bacillus subtilis and Shenqu (a type of fermented wheat) to conduct synergistic aerobic fermentation. Through metabolic activity, it rapidly degrades crude fiber in bran, dietary fiber in fruit peels, and bound macromolecular polysaccharides in Shenqu and malt, reducing the structural complexity of the raw materials. Simultaneously, it secretes extracellular enzymes such as amylase, saccharifying enzymes, and cellulase to initially enzymatically hydrolyze starch and polysaccharide nutrients. This process not only significantly improves the bioavailability of the materials but also, through rapid microbial metabolism, raises the temperature and softens the materials, creating ideal physical conditions, a rich enzyme base, and an active microbial community for the subsequent second-stage anaerobic fermentation.

[0035] In this invention, after the first stage of aerobic fermentation is completed, the second stage of anaerobic fermentation is carried out. The temperature of the anaerobic fermentation is preferably 25~35℃, more preferably 28~32℃, and even more preferably 30℃. The time of the anaerobic fermentation is preferably 96~144h, more preferably 108~132h, and even more preferably 120h.

[0036] This invention utilizes anaerobic fermentation, primarily by lactic acid bacteria and yeast, to further degrade incompletely decomposed macromolecular nutrients, transforming them into smaller peptides, organic acids, monosaccharides, and other nutrients more easily digested and absorbed by aquatic animals. Simultaneously, under a low-oxygen fermentation environment, it synergistically promotes the dissolution and enrichment of core active ingredients such as saponins, polysaccharides, and flavonoids from traditional Chinese medicine raw materials like Astragalus membranaceus, Dioscorea opposita, and Poria cocos, while also activating active ingredients in Shenqu (a type of fermented medicinal material) related to aquatic animal digestion and immunity. The lactic acid and acetic acid produced by the anaerobic bacteria effectively lower the pH of the material, inhibiting the growth of harmful bacteria. Furthermore, the organic acids synergize with the enzymes secreted during the aerobic fermentation stage, further enhancing the overall enzymatic hydrolysis level of the material. Simultaneously, during fermentation, the endogenous and exogenous anaerobic bacteria in Shenqu achieve synergistic proliferation, constructing a stable beneficial microbial system. This provides a foundation for optimizing the intestinal microecological structure and enhancing the immunity of aquatic animals during subsequent feeding.

[0037] This invention utilizes a two-stage fermentation process to fully leverage the synergistic effect of substrate degradation and functional microbial communities, significantly improving the final body weight, weight gain rate, specific growth rate, and average survival rate of Litopenaeus vannamei, enhancing the disease resistance of Litopenaeus vannamei, and reducing mortality after viral challenge.

[0038] The present invention also provides the application of the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method in the preparation of products that promote the growth of aquatic animals and / or prevent aquatic animal diseases.

[0039] In this invention, the species of aquatic animals are not specifically limited, but are preferably including whiteleg shrimp, tiger prawn, or giant freshwater prawn.

[0040] In this invention, the aquatic animal diseases preferably include bacterial diseases; the bacterial diseases preferably include aquatic animal diseases caused by Vibrio parahaemolyticus.

[0041] The present invention also provides a method for preventing diseases in aquatic animals, the method comprising: mixing the traditional Chinese medicine composition or the traditional Chinese medicine composition prepared by the preparation method with feed and feeding it to aquatic animals.

[0042] In this invention, the amount of the traditional Chinese medicine composition added is preferably 0.5% to 3% of the feed mass, more preferably 1% to 2%, and even more preferably 1.5%.

[0043] In this invention, the mixing method is not particularly limited. In some embodiments, the traditional Chinese medicine composition is mixed with feed in a certain proportion, water is added to make the traditional Chinese medicine composition fully adhere to the feed, and then it is air-dried at room temperature and sealed for storage.

[0044] The aquaculture method of this invention can effectively improve the growth performance and disease resistance of aquatic animals.

[0045] 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.

[0046] Unless otherwise specified, the following embodiments are all conventional methods.

[0047] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.

[0048] The Bacillus subtilis inoculant used in the following examples was purchased from Jinan Chaoyixing Chemical Co., Ltd., with a viable count of 100 billion CFU / g. The Bacillus subtilis bacterial solution used was obtained by dissolving the Bacillus subtilis inoculant in water to obtain a Bacillus subtilis bacterial solution with the corresponding viable bacterial concentration.

[0049] Example 1 A traditional Chinese medicine composition for preventing diseases in aquatic animals, the composition being made from the following raw materials: 15 parts Astragalus membranaceus, 10 parts Dioscorea opposita, 10 parts Poria cocos, 20 parts Massa fermentata, 10 parts Hordeum vulgare, 15 parts Litchi chinensis peel, 15 parts wheat bran, and 5 parts Glycyrrhiza uralensis.

[0050] The preparation method of the traditional Chinese medicine composition includes the following steps: weighing each raw material according to the weight parts, pulverizing, sieving through a 60-mesh sieve, mixing to obtain a mixture; adding water to the mixture to adjust the moisture content to 45%, and inoculating with Bacillus subtilis bacterial solution (1×10⁻⁶). 9 The inoculum of Bacillus subtilis (cfu / mL) was 0.5% (v / w, mL / g) of the dry weight of the mixture. The first stage of aerobic fermentation was carried out at 35℃, with the material being turned over every 10 hours during the aerobic fermentation process, and the fermentation time was 72 hours. After the first stage of aerobic fermentation was completed, the second stage of anaerobic fermentation was carried out at 30℃ for 120 hours. After the second stage of anaerobic fermentation was completed, the mixture was dried to obtain the traditional Chinese medicine composition.

[0051] Example 2 A traditional Chinese medicine composition for preventing diseases in aquatic animals, the composition being made from the following raw materials: 10 parts Astragalus membranaceus, 5 parts Dioscorea opposita, 5 parts Poria cocos, 10 parts Massa fermentata, 5 parts malt, 10 parts pineapple peel, 10 parts wheat bran, and 2 parts Glycyrrhiza uralensis.

[0052] The preparation method of the traditional Chinese medicine composition includes the following steps: weighing each raw material according to the weight parts, pulverizing, sieving through a 40-mesh sieve, mixing to obtain a mixture; adding water to the mixture to adjust the moisture content to 40%, and inoculating with Bacillus subtilis bacterial solution (1×10⁻⁶). 8 The inoculum of Bacillus subtilis was 0.3% (v / w, mL / g) of the dry weight of the mixture. The first stage of aerobic fermentation was carried out at 30℃, with the material being turned over every 12 hours during the aerobic fermentation process, and the fermentation time was 80 hours. After the first stage of aerobic fermentation was completed, the second stage of anaerobic fermentation was carried out at 25℃ for 144 hours. After the second stage of anaerobic fermentation was completed, the mixture was dried to obtain the traditional Chinese medicine composition.

[0053] Example 3 A traditional Chinese medicine composition for preventing diseases in aquatic animals, the composition being made from the following raw materials: 20 parts Astragalus membranaceus, 15 parts Dioscorea opposita, 15 parts Poria cocos, 30 parts Massa fermentata, 15 parts malt, 10 parts banana peel, 10 parts dragon fruit peel, 20 parts wheat bran, and 8 parts licorice.

[0054] The preparation method of the traditional Chinese medicine composition includes the following steps: weighing each raw material according to the weight parts, pulverizing, sieving through an 80-mesh sieve, mixing to obtain a mixture; adding water to the mixture to adjust the moisture content to 50%, and inoculating with Bacillus subtilis bacterial solution (1×10⁻⁶). 10The inoculum of Bacillus subtilis was 0.8% (v / w, mL / g) of the dry weight of the mixture. The first stage of aerobic fermentation was carried out at 40℃, with the material being turned over every 8 hours during the aerobic fermentation process, and the fermentation time was 60 hours. After the first stage of aerobic fermentation was completed, the second stage of anaerobic fermentation was carried out at 35℃ for 96 hours. After the second stage of anaerobic fermentation was completed, the mixture was dried to obtain the traditional Chinese medicine composition.

[0055] Example 4 A method for preventing diseases in aquatic animals, wherein the method comprises: mixing the traditional Chinese medicine composition of Example 1 with feed and feeding it to aquatic animals; the amount of the traditional Chinese medicine composition added is 1.5% of the feed mass.

[0056] Example 5 A method for preventing diseases in aquatic animals, wherein the method comprises: mixing the traditional Chinese medicine composition of Example 2 with feed and feeding it to aquatic animals; the amount of the traditional Chinese medicine composition added is 3% of the feed mass.

[0057] Example 6 A method for preventing diseases in aquatic animals, wherein the method comprises: mixing the traditional Chinese medicine composition of Example 3 with feed and feeding it to aquatic animals; the amount of the traditional Chinese medicine composition added is 0.5% of the feed mass.

[0058] Comparative Example 1 A traditional Chinese medicine composition for preventing diseases in aquatic animals, the only difference from Example 1 is that an equal amount of inactivated Shenqu is used to replace Shenqu; The preparation method of the inactivated Shenqu is as follows: sterilize with high-pressure steam at 121℃ and 0.1MPa for 30 minutes, dry at 60℃ to constant weight, and pulverize through a 60-mesh sieve to obtain inactivated Shenqu.

[0059] Comparative Example 2 A traditional Chinese medicine composition for preventing diseases in aquatic animals, differing from Example 1 only in that the preparation method of the traditional Chinese medicine composition is as follows: Weigh each raw material according to weight parts, pulverize, sieve through a 60-mesh sieve, mix, and obtain a mixture; add water to the mixture to adjust the moisture content to 45%, and inoculate with Bacillus subtilis bacterial solution (1×10⁻⁶). 9 The inoculum of Bacillus subtilis was 0.5% (v / w, mL / g) of the dry weight of the mixture. Aerobic fermentation was carried out at 35℃. During the aerobic fermentation, the material was turned over every 10 hours, and the fermentation time was 72 hours. After the aerobic fermentation was completed, the mixture was dried to obtain the traditional Chinese medicine composition.

[0060] Comparative Example 3 A traditional Chinese medicine composition for preventing diseases in aquatic animals, the only difference from Example 1 is that the preparation method of the traditional Chinese medicine composition is as follows: weigh each raw material according to the weight parts, crush it, sieve it through a 60-mesh sieve, mix it to obtain a mixture; add water to the mixture to adjust the moisture content to 45%, carry out anaerobic fermentation at a fermentation temperature of 30°C for 120 hours, after the anaerobic fermentation is completed, dry it to obtain the traditional Chinese medicine composition.

[0061] Comparative Example 4 A traditional Chinese medicine composition for preventing diseases in aquatic animals, differing from Comparative Example 1 only in that the preparation method of the traditional Chinese medicine composition includes the following steps: weighing each raw material according to weight parts, pulverizing, sieving through a 60-mesh sieve, mixing to obtain a mixture; adding water to the mixture to adjust the moisture content to 45%, and inoculating with Bacillus subtilis bacterial solution (1×10⁻⁶). 9 The inoculum of Bacillus subtilis was 0.5% (v / w, mL / g) of the dry weight of the mixture. The first stage of aerobic fermentation was carried out at 35℃, with the material being turned over every 10 hours for a total fermentation time of 72 hours. After the first stage of aerobic fermentation, 0.5% of the compound bacterial solution was added (where the ratio of live Saccharomyces cerevisiae to Lactobacillus plantarum was 1:1, and the total live count was ≥1×10⁻⁶). 8 The second stage of anaerobic fermentation was carried out at 30℃ for 120 hours (CFU / mL). After the second stage of anaerobic fermentation was completed, the mixture was dried to obtain the traditional Chinese medicine composition.

[0062] Experimental Example 1 1. Experimental Methods Healthy and vigorous Pacific white shrimp with an initial weight of 3.0 ± 0.2 g were selected for the experiment and were acclimatized indoors for 7 days before the experiment. The shrimp were randomly divided into 6 groups, with 3 replicates per group. Each replicate was placed in a 500L culture tank, and 30 shrimp were stocked in each tank. The experimental groups are as follows: (1) Blank control group: fed basic feed without any added preparations; (2) Experimental group: fed with basic feed + 1.5% of the traditional Chinese medicine composition of Example 1; (3) Control group 1: fed with basic feed + 1.5% of the traditional Chinese medicine composition of control group 1; (4) Control group 2: fed with basic feed + 1.5% of the traditional Chinese medicine composition of control group 2; (5) Control group 3: fed with basic feed + 1.5% of the traditional Chinese medicine composition of control group 3; (6) Control group 4: fed with basic feed + 1.5% of the Chinese herbal composition of control group 4.

[0063] Each group mixed the corresponding Chinese herbal medicine composition with the basic feed in a certain proportion, added a small amount of water to make the Chinese herbal medicine composition fully adhere to the basic feed, air-dried at room temperature and then sealed and stored to obtain the feed for each group.

[0064] Each breeding tank was kept under the same environmental conditions, fed 3 times a day, with a daily feed amount of about 6% to 8% of the body weight of the whiteleg shrimp, and about 30% of the water was changed daily. The experimental period was 28 days.

[0065] 2. Indicator Measurement Before and after the experiment, the weight of each group of whiteleg shrimp was measured and the number of surviving shrimp was recorded. The indicators were calculated according to the following formulas, and the results are shown in Table 1.

[0066] Weight gain rate = (average final weight – average initial weight) / average initial weight × 100% Specific growth rate = (Ln average final body weight – Ln average initial body weight) / 28 days × 100% Survival rate = (Number of terminal tails / Number of initial tails) × 100% Table 1. Results of index measurement for each group

[0067] Note: Different lowercase letters in the same column indicate significant differences (P<0.05).

[0068] As shown in Table 1, the experimental group, using active Shenqu combined with a two-stage fermentation process, fully leveraged the synergistic effect of substrate degradation and functional microbial communities, significantly improving the final body weight, weight gain rate, specific growth rate, and average survival rate of Litopenaeus vannamei. Control group 1, using inactivated Shenqu without exogenous microbial supplementation, despite undergoing two-stage fermentation, showed only slightly better growth and survival performance than the control group due to the lack of endogenous functional microbial communities and the inability to generate effective active ingredients caused by the inactivation of Shenqu. This highlights that active Shenqu and endogenous microbial communities are the core of its efficacy. Control groups 2 and 3, using only aerobic or anaerobic fermentation, showed significantly better performance than the blank control group and control group 1 due to insufficient fermentation, inadequate substrate degradation, and insufficient release of effective ingredients, but still far inferior to the experimental group, demonstrating the necessity of a complete two-stage fermentation process for improving efficacy. Although the Shenqu in control group 4 was inactivated, the addition of exogenous functional microorganisms partially compensated for the disadvantage of the lack of microorganisms. Its performance was significantly better than the blank control group, control group 1 and the single-stage fermentation group, but still significantly lower than the experimental group. This indicates that although exogenous microorganisms can improve the effect, they cannot completely replace the synergistic effect of active Shenqu and endogenous microorganisms in the two-stage fermentation.

[0069] Experimental Example 2 A pathogenic Vibrio parahaemolyticus strain was isolated and identified from diseased Litopenaeus vannamei. After large-scale culture, the supernatant was removed by centrifugation, and the culture was resuspended in sterile physiological saline to obtain a concentration of 1×10⁻⁶. 7CFU / mL bacterial culture.

[0070] In Experiment 1, after the experiment was completed, 15 healthy and vigorous shrimp were randomly selected from each parallel experiment and injected intramuscularly with 0.15 mL of the above-mentioned Vibrio parahaemolyticus bacterial solution. After the injection, the shrimp were placed in a new culture tank and continued to be raised under the same conditions. During this period, the shrimp were fed a basic feed and observed for 7 consecutive days. The number of dead shrimp on the 7th day was recorded, and the cumulative mortality rate was calculated. The results are shown in Table 2.

[0071] Table 2. Cumulative Mortality Rate on Day 7 of Infection Challenge

[0072] As shown in Table 2, the experimental group, using active Shenqu combined with a two-stage fermentation process, exhibited the best disease resistance in shrimp, with a significantly lower cumulative mortality rate than other groups, demonstrating that this treatment effectively enhances shrimp's resistance to pathogens. Control group 1, using inactivated Shenqu, had a significantly higher cumulative mortality rate than all groups except the blank control group, indicating that active Shenqu is the core foundation for improving shrimp disease resistance. Control groups 2 and 3, using only aerobic or anaerobic fermentation processes, showed no significant difference in cumulative mortality rates, both significantly higher than control group 4 and the experimental group, indicating that single-stage fermentation is insufficient to fully leverage the disease-regulating effect of feed. Although control group 4 used inactivated Shenqu, the addition of exogenous functional bacteria compensated for some of the deficiency in endogenous bacteria; however, its cumulative mortality rate was still significantly higher than the experimental group, indicating that while exogenous functional bacteria can improve shrimp disease resistance to some extent, they cannot completely replace the synergistic effect of active Shenqu and the two-stage fermentation process.

[0073] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A traditional Chinese medicine composition for preventing diseases in aquatic animals, characterized in that, The traditional Chinese medicine composition comprises the following raw materials in parts by weight: 10-20 parts of Astragalus membranaceus, 5-15 parts of Dioscorea opposita, 5-15 parts of Poria cocos, 10-30 parts of Medicated Leaven, 5-15 parts of Malt, 10-20 parts of Pericarpium Citri Reticulatae, 10-20 parts of Wheat Bran, and 2-8 parts of Glycyrrhiza uralensis.

2. The traditional Chinese medicine composition according to claim 1, characterized in that, The peel includes any one or more of the following: lychee peel, pineapple peel, banana peel, and dragon fruit peel.

3. The method for preparing the traditional Chinese medicine composition according to any one of claims 1 to 2, characterized in that, The preparation method includes: weighing each raw material according to the weight parts, crushing and sieving, mixing to obtain a mixture; inoculating the mixture with Bacillus subtilis bacterial solution, and sequentially carrying out a first stage of aerobic fermentation and a second stage of anaerobic fermentation; after the second stage of anaerobic fermentation is completed, drying to obtain a traditional Chinese medicine composition.

4. The preparation method according to claim 3, characterized in that, Before adding Bacillus subtilis bacterial solution to the mixture, the moisture content is adjusted to 40%~50%.

5. The preparation method according to claim 3, characterized in that, The viable cell concentration of the Bacillus subtilis bacterial suspension was 1×10⁻⁶. 8 ~1×10 10 cfu / mL; the inoculation amount of the Bacillus subtilis bacterial solution is 0.3%~0.8% of the dry weight of the mixture.

6. The preparation method according to claim 3, characterized in that, The conditions for aerobic fermentation include: fermentation temperature of 30~40℃ and time of 60~80h.

7. The preparation method according to claim 3, characterized in that, The conditions for anaerobic fermentation include: fermentation temperature of 25~35℃ and time of 96~144h.

8. The use of the traditional Chinese medicine composition according to any one of claims 1 to 2 or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 3 to 7 in the preparation of products that promote the growth of aquatic animals and / or prevent aquatic animal diseases.

9. A method for preventing diseases in aquatic animals, characterized in that, The breeding method includes: mixing the traditional Chinese medicine composition according to any one of claims 1 to 2 or the traditional Chinese medicine composition prepared by the preparation method according to any one of claims 3 to 7 with feed, and feeding it to aquatic animals.

10. The aquaculture method according to claim 9, characterized in that, The amount of the traditional Chinese medicine composition added is 0.5% to 3% of the feed mass.

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