Compound fermented Chinese herbal medicine feed for enhancing immunity of groupers and application of compound fermented Chinese herbal medicine feed

Abstract

The invention relates to the related technical field of fish feeds, in particular to a compound fermented Chinese herbal medicine feed for enhancing the immunity of groupers and application of the compound fermented Chinese herbal medicine feed, and the feed is prepared from the following components in percentage by weight: 85%-92% of basic feed raw materials, 5%-12% of compound fermented Chinese herbal medicine additives and 2%-5% of functional auxiliary agents. The core of the additive is that the additive consists of astragalus membranaceus, rheum officinale, andrographis paniculata and liquorice according to a specific ratio of (3-4): (2-3): (1-2): (0.5-1), and is prepared by fermenting the astragalus membranaceus, the rheum officinale, the andrographis paniculata and the liquorice with a compound strain of lactobacillus plantarum and bacillus subtilis at 30-37 DEG C for 36-60 hours. According to the feed, the activity of serum lysozyme and superoxide dismutase of groupers can be remarkably improved, the non-specific immunity is enhanced, the survival rate of the groupers infected by vibrio alginolyticus is not lower than 85% in a toxicity attacking experiment, and vibriosis and cryptocaryon irritans are effectively prevented.

Description

Technical Field

[0001] This invention relates to the field of fish feed technology, and in particular to a compound fermented herbal feed for enhancing the immunity of grouper and its application. Background Technology

[0002] Grouper is an important commercially important marine aquaculture fish worldwide, prized for its delicious and nutritious flesh and strong market demand, leading to a vast industry chain along my country's southeastern coast. However, grouper is highly susceptible to various diseases during intensive aquaculture, such as viral neuronecrosis (VNN), vibriosis, cryptocaryonosis, and trypanosomiasis. These diseases often result in weakened immunity, stunted growth, and even mass mortality, causing significant economic losses for fish farmers. Currently, farmers rely heavily on feeding fresh miscellaneous fish or using chemical drugs for disease prevention and control. However, fresh miscellaneous fish can easily carry pathogens and pollute water, while chemical drugs pose risks such as drug residues, environmental pollution, and the development of drug resistance in pathogens. Therefore, developing nutritional strategies that can fundamentally enhance the grouper's own immunity has become an urgent need for the healthy development of the industry.

[0003] A search revealed that patent CN102090537A discloses a powdered compound feed for grouper. Its core components are conventional raw materials such as steam-dried fishmeal (up to 66%) and pregelatinized starch, supplemented with a microecological agent (Bacillus subtilis) and an attractant (betaine). This technical solution primarily focuses on providing basic nutrition, reducing feed costs, and improving palatability. However, its shortcomings are: 1) its formula is essentially a combination of conventional nutrients, lacking functional active ingredients that can specifically activate the grouper's immune system; 2) the microecological agent used is added directly without undergoing synergistic fermentation with specific functional components, resulting in limited probiotic effects and failing to achieve synergistic immune regulation. Therefore, this feed cannot effectively address complex disease challenges, especially in preventing specific diseases such as vibriosis and cryptocaryonosis.

[0004] In light of this, in-depth research into the aforementioned issues led to the creation of this case. Summary of the Invention

[0005] The purpose of this invention is to provide a compound fermented herbal feed for grouper that enhances the immunity of grouper and its application, in order to solve the problems mentioned in the background art, such as the inability of traditional feed to effectively cope with complex disease challenges, especially the poor effect in preventing specific diseases such as vibrio and cryptocaryonosis.

[0006] To achieve the above objectives, the present invention provides the following technical solution: a compound fermented herbal feed for enhancing the immunity of grouper, comprising the following components by weight percentage:

[0007] Basic feed ingredients: 85%–92%;

[0008] Compound fermented Chinese herbal medicine additives: 5%–12%;

[0009] Functional adjuvants 2%–5%;

[0010] The compound fermented herbal additive is a compound composed of Astragalus membranaceus, Rheum palmatum, Andrographis paniculata and Glycyrrhiza uralensis in a weight ratio of (3-4):(2-3):(1-2):(0.5-1), which is fermented by a compound strain of Lactobacillus plantarum and Bacillus subtilis at 30-37°C for 36-60 hours.

[0011] Preferably, the preparation method of the compound fermented herbal medicine additive includes the following steps:

[0012] a) Grind Astragalus membranaceus, Rheum palmatum, Andrographis paniculata, and Glycyrrhiza uralensis into 60-80 mesh respectively, and mix them evenly according to the weight ratio described above;

[0013] b) Mix the mixed Chinese herbal medicine powder with the culture medium at a mass ratio of 1:(4-6), wherein the culture medium is prepared by brown sugar, soybean meal and water at a mass ratio of 1:5:10;

[0014] c) Inoculate the mixture from step b) with a compound microbial strain for fermentation, wherein the compound microbial strain consists of Lactobacillus plantarum and Bacillus subtilis, and the ratio of viable cells (Lactobacillus plantarum: Bacillus subtilis) is (1-2):1, and the total inoculation amount is 3%-5% of the total mass of the mixture;

[0015] d) Ferment at 30–37°C for 36–60 hours;

[0016] e) After fermentation, the fermentation product is dried at a low temperature of 50-60℃, pulverized and passed through a 60-mesh sieve to obtain the compound fermented Chinese herbal medicine additive.

[0017] Preferably, the basic feed ingredients consist of the following components by weight percentage: 40%–50% fishmeal, 15%–20% fermented soybean meal, 20%–30% high-gluten flour, and 2%–4% brewer's yeast.

[0018] Preferably, the functional adjuvant includes vitamin C in a content of 0.1% to 0.3%, choline chloride in a content of 0.2% to 0.5%, lecithin in a content of 1% to 2%, and astaxanthin in a content of 0.05% to 0.15%.

[0019] Application of a compound fermented herbal feed that enhances the immunity of grouper in the preparation of feed for the prevention and / or adjunctive treatment of vibriosis and cryptocaryonosis in grouper.

[0020] The preferred feeding amount is 3% to 5% of the grouper's body weight, fed 2 to 3 times a day.

[0021] The preferred application can significantly increase the activity of lysozyme and superoxide dismutase (SOD) in grouper serum.

[0022] Preferably, the grouper fed with the aforementioned feed has a survival rate of no less than 85% after being artificially infected with Vibrio alginolyticus.

[0023] The medicinal effects of the above-mentioned Chinese herbal medicines are as follows:

[0024] Astragalus: Its rich astragalus polysaccharides and other active ingredients can fundamentally stimulate and strengthen the non-specific immune system of grouper. By significantly improving key indicators such as serum lysozyme activity, it establishes a strong basic immune defense for the fish and plays a decisive role in "tonifying qi and consolidating the exterior".

[0025] Rhubarb: Its main active ingredient, anthraquinone compounds, can precisely inhibit the growth and reproduction of common pathogenic bacteria such as Vibrio alginolyticus, effectively preventing and treating serious vibriosis. It complements the immune-enhancing effect of Astragalus membranaceus, together forming a synergistic system of "prevention is better than cure".

[0026] Andrographis paniculata: Its unique andrographolide can not only effectively combat viral threats such as viral neuronecrosis, but also cleverly regulate the intensity of the immune response, prevent the immune system from being over-activated and causing damage to the body, thereby ensuring a more balanced and safe immune enhancement process.

[0027] Licorice: Glycyrrhizic acid can not only alleviate the irritation of rhubarb and improve the overall palatability of feed, but also synergistically enhance the anti-inflammatory effects of other medicinal materials and significantly improve the tolerance of grouper to water quality fluctuations and high temperatures, ensuring that the entire compound exerts a stable and comprehensive effect.

[0028] Compared with the prior art, the beneficial effects of the present invention are:

[0029] 1. This invention utilizes a unique compound fermentation process to fully release and transform the effective components of traditional Chinese medicine, systematically and significantly enhancing the non-specific immune level of grouper. A specific ratio of Astragalus membranaceus, rhubarb, Andrographis paniculata, and licorice, after synergistic fermentation by Lactobacillus plantarum and Bacillus subtilis, greatly improves the bioavailability of its immunomodulatory substances (such as polysaccharides and flavonoids). After ingestion, these highly effective components directly act on the grouper's immune system, significantly increasing the activity of key immune indicators such as serum lysozyme and superoxide dismutase (SOD). This means that the grouper's ability to resist pathogen invasion and clear oxidative damage is fundamentally enhanced, laying a solid physiological foundation for disease resistance.

[0030] 2. This feed can translate enhanced immunity into superior prevention against specific major diseases, directly reducing mortality during the aquaculture process and resulting in significant economic benefits. Grouper fed this feed long-term exhibit strong resistance to Vibrio alginolyticus infection and Cryptocaryon irritans infestation, consistently improving survival rates far superior to grouper fed ordinary feed. This targeted and effective protection effectively reduces sudden mortality caused by disease outbreaks, ensuring high survival rates and final yields, providing farmers with a stable and reliable risk control solution.

[0031] 3. This invention realizes a safe, environmentally friendly, and highly efficient healthy aquaculture model that meets the requirements of sustainable development in aquaculture. Since the core ingredients are natural Chinese herbal medicines and probiotics, the use of antibiotics and chemical drugs is completely avoided, thus eliminating the problem of drug residues and producing safer aquatic products. At the same time, healthy fish have a higher feed conversion and utilization rate, and less nitrogen and phosphorus waste is discharged into the water. In addition, their enhanced stress resistance helps fish to cope smoothly with water quality fluctuations and high temperature stress, jointly maintaining a good aquaculture environment and forming a virtuous cycle chain from internal immunity to external environment. Attached Figure Description

[0032] Figure 1 This is a schematic diagram of the complete process of the present invention from raw material preparation to final application. Detailed Implementation

[0033] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0034] like Figure 1 As shown, the present invention provides the following technical solution: a compound fermented herbal feed for enhancing the immunity of grouper, comprising the following components by weight percentage:

[0035] Basic feed ingredients: 85%–92%;

[0036] Compound fermented Chinese herbal medicine additives: 5%–12%;

[0037] Functional adjuvants 2%–5%;

[0038] The compound fermented herbal additive is a compound composed of Astragalus membranaceus, Rheum palmatum, Andrographis paniculata and Glycyrrhiza uralensis in a weight ratio of (3-4):(2-3):(1-2):(0.5-1), which is fermented by a compound strain of Lactobacillus plantarum and Bacillus subtilis at 30-37°C for 36-60 hours.

[0039] Furthermore, the preparation method of the compound fermented herbal medicine additive includes the following steps:

[0040] a) Grind Astragalus membranaceus, Rheum palmatum, Andrographis paniculata, and Glycyrrhiza uralensis into 60-80 mesh respectively, and mix them evenly according to the weight ratio described above;

[0041] b) Mix the mixed Chinese herbal medicine powder with the culture medium at a mass ratio of 1:(4-6), wherein the culture medium is prepared by brown sugar, soybean meal and water at a mass ratio of 1:5:10;

[0042] c) Inoculate the mixture from step b) with a compound microbial strain for fermentation, wherein the compound microbial strain consists of Lactobacillus plantarum and Bacillus subtilis, and the ratio of viable cells (Lactobacillus plantarum: Bacillus subtilis) is (1-2):1, and the total inoculation amount is 3%-5% of the total mass of the mixture;

[0043] d) Ferment at 30–37°C for 36–60 hours;

[0044] e) After fermentation, the fermentation product is dried at a low temperature of 50-60℃, pulverized and passed through a 60-mesh sieve to obtain the compound fermented Chinese herbal medicine additive.

[0045] Furthermore, the basic feed ingredients consist of the following components by weight percentage: 40%–50% fishmeal, 15%–20% fermented soybean meal, 20%–30% high-gluten flour, and 2%–4% brewer's yeast.

[0046] Furthermore, the functional adjuvant includes vitamin C in a content of 0.1% to 0.3%, choline chloride in a content of 0.2% to 0.5%, lecithin in a content of 1% to 2%, and astaxanthin in a content of 0.05% to 0.15%.

[0047] Example: Preparation, breeding effects and disease resistance evaluation of compound fermented Chinese herbal medicine feed

[0048] Example 1: Preparation of Compound Fermented Traditional Chinese Medicine Additive

[0049] Ingredients: 30 kg of dried Astragalus membranaceus, 20 kg of Rheum palmatum, 15 kg of Andrographis paniculata, and 5 kg of Glycyrrhiza uralensis.

[0050] Pretreatment: Grind the above-mentioned Chinese herbal medicines separately using a grinder, pass them through a 60-mesh sieve, and mix them evenly.

[0051] Culture medium preparation: Take 1 kg of brown sugar, 5 kg of soybean meal and 10 kg of purified water, stir and mix evenly to make culture medium.

[0052] Inoculation and Fermentation: Mix 12 kg of the mixed herbal powder with all the above-mentioned culture medium (approximately 16 kg) and add it to the fermenter. Then inoculate with a compound bacterial strain (Lactobacillus plantarum freeze-dried powder: Bacillus subtilis freeze-dried powder = 1.5:1 (live count ratio)), with a total inoculation amount of 4% of the total mass of the fermentation system (i.e., 1.12 kg). Ferment at 35°C with aeration and stirring for 48 hours.

[0053] Post-processing: After fermentation, the product is dried at 55℃ until the moisture content is less than 10%, then pulverized again and passed through a 60-mesh sieve to obtain the compound fermented Chinese herbal medicine additive, which is then sealed and packaged for later use.

[0054] Example 2: Formulation of Compound Fermented Herbal Feed

[0055] Formula: Prepare 1000 kg of experimental feed according to the following formula:

[0056] The basic feed ingredients account for 88.5%, specifically including 45% fishmeal (450 kg), 18% fermented soybean meal (180 kg), 24% high-gluten flour (240 kg), and 1.5% brewer's yeast (15 kg). 8% (80 kg) of the compound fermented herbal additive prepared in Example 1 is added. Functional adjuvants account for 3.5%, including 0.2% vitamin C (2 kg), 0.3% choline chloride (3 kg), 1.5% lecithin (15 kg), 0.5% astaxanthin (10% content, actually 5 kg), and 0.5% betaine (5 kg). Finally, 0.5% mineral premix (5 kg) is added.

[0057] Processing technology: All the above raw materials are put into a twin-shaft paddle mixer and mixed evenly. Then, they are fed into a conditioner and steam is introduced for conditioning. Finally, they are pelleted at 85°C (particle size 3.0 mm) by a twin-screw pellet mill. After cooling and packaging, they become finished feed.

[0058] Example 3: Aquaculture Experiment and Immune Index Measurement

[0059] Experimental design: 600 healthy juvenile tiger grouper with an average weight of (50.5±2.0) g were randomly divided into 2 groups, with 3 replicates in each group and 100 fish in each replicate. The experiment was conducted in an indoor cement pond (5m5m1.5m) for 60 days.

[0060] Control group: fed with a commercially available brand of grouper compound feed (66% fish meal, 10% soybean meal, 18% wheat flour, etc.).

[0061] Experimental group: fed with the feed of the present invention prepared in Example 2.

[0062] Feeding management: Feed twice a day (08:00 and 17:00), with the amount of feed being 3%-5% of the fish's body weight, and the specific amount should be adjusted slightly according to the feeding situation and water temperature.

[0063] Results and Analysis:

[0064] Growth Performance: After the experiment, growth performance and immune indicators were evaluated. In terms of growth performance, the experimental group's average final weight reached 245.7 g, significantly higher than the control group's 218.5 g. The experimental group's weight gain rate was 385.6%, significantly higher than the control group's 334.4%. Simultaneously, the experimental group's feed conversion ratio was 1.42, significantly better than the control group's 1.58. Regarding immune indicators, the experimental group's serum lysozyme activity reached 168.9 units per milliliter, and superoxide dismutase activity reached 108.7 units per milliliter. Both of these key immune indicators were significantly higher than the control group (lysozyme 125.6 U / mL, superoxide dismutase 85.4 U / mL).

[0065] Example 4: Challenge Test (Evaluation of Disease Resistance)

[0066] Experimental method: After the aquaculture experiment, 30 fish were randomly selected from each replicate (i.e., 90 fish per group) for Vibrio alginolyticus challenge test.

[0067] Challenge procedure: The pathogenic Vibrio alginolyticus strain was revived and cultured, and the bacterial concentration was adjusted to 1×10^7 CFU / mL with physiological saline. 0.1 mL of the bacterial solution was injected intraperitoneally into each fish. The fish were observed for 7 consecutive days after challenge, and mortality was recorded. Dead fish were promptly removed and the pathogen was re-isolated and confirmed.

[0068] Results and Analysis: After a 7-day observation period following the viral challenge, the results showed that the cumulative mortality rate of the experimental group (15.6%) was significantly lower than that of the control group (33.3%), with a relative protection rate as high as 76.7%. This strongly demonstrates that the feed of this invention can significantly enhance the resistance of grouper to major bacterial diseases.

[0069] Although embodiments of the invention have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and alterations can be made to these embodiments without departing from the principles and spirit of the invention, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A compound fermented herbal feed for enhancing the immunity of grouper, characterized in that, It consists of the following components by weight percentage: Basic feed ingredients: 85%–92%; Compound fermented Chinese herbal medicine additives: 5%–12%; Functional adjuvants 2%–5%; The compound fermented herbal additive is a compound composed of Astragalus membranaceus, Rheum palmatum, Andrographis paniculata and Glycyrrhiza uralensis in a weight ratio of (3-4):(2-3):(1-2):(0.5-1), which is fermented by a compound strain of Lactobacillus plantarum and Bacillus subtilis at 30-37°C for 36-60 hours.

2. The compound fermented herbal feed for enhancing the immunity of grouper according to claim 1, characterized in that, The preparation method of the compound fermented herbal medicine additive includes the following steps: a) Grind Astragalus membranaceus, Rheum palmatum, Andrographis paniculata, and Glycyrrhiza uralensis into 60-80 mesh respectively, and mix them evenly according to the weight ratio described above; b) Mix the mixed Chinese herbal medicine powder with the culture medium at a mass ratio of 1:(4-6), wherein the culture medium is prepared by brown sugar, soybean meal and water at a mass ratio of 1:5:10; c) Inoculate the mixture from step b) with a compound microbial strain for fermentation, wherein the compound microbial strain consists of Lactobacillus plantarum and Bacillus subtilis, and the ratio of viable cells (Lactobacillus plantarum: Bacillus subtilis) is (1-2):1, and the total inoculation amount is 3%-5% of the total mass of the mixture; d) Ferment at 30–37°C for 36–60 hours; e) After fermentation, the fermentation product is dried at a low temperature of 50-60℃, pulverized and passed through a 60-mesh sieve to obtain the compound fermented Chinese herbal medicine additive.

3. The compound fermented herbal feed for enhancing the immunity of grouper according to claim 1, characterized in that, The basic feed ingredients consist of the following components by weight percentage: 40%–50% fishmeal, 15%–20% fermented soybean meal, 20%–30% high-gluten flour, and 2%–4% brewer's yeast.

4. The compound fermented herbal feed for enhancing the immunity of grouper according to claim 1, characterized in that, The functional adjuvants include vitamin C, with a content of 0.1% to 0.3%; choline chloride, with a content of 0.2% to 0.5%; lecithin, with a content of 1% to 2%; and astaxanthin, with a content of 0.05% to 0.15%.

5. The use of a compound fermented herbal feed for enhancing the immunity of grouper as described in any one of claims 1-4 in the preparation of feed for the prevention and / or adjunctive treatment of vibriosis and cryptocaryonosis in grouper.

6. The application according to claim 5, characterized in that, The feed should be given at a rate of 3% to 5% of the grouper's body weight, 2 to 3 times a day.

7. The application according to claim 5, characterized in that, The application significantly increases the activity of lysozyme and superoxide dismutase (SOD) in grouper serum.

8. The application according to claim 5, characterized in that, The survival rate of grouper fed with the aforementioned feed after artificial infection with Vibrio alginolyticus is no less than 85%.

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