Preparation method of a compound microecological preparation for penaeus vannamei cultivation

Abstract

The application provides a preparation method of a white shrimp (Penaeus vannamei) breeding compound microecological preparation, which comprises the following steps: S1, strain selection; S2, enrichment culture; S3, preparation of a suspension; and S4, blending, and the like. Compound effective bacteria with a probiotic effect and strong environmental adaptability are screened, inoculated into an activated culture medium, and a microbial community formed by the inoculation is helpful to the formation of a benign circulating ecological system. Through enrichment culture, the growth speed and metabolic activity of the strain are improved, the adaptability of the strain is promoted, the diversity of the strain is increased, and the enzyme activity of the strain is improved. Antibacterial peptides and regulators are added to inhibit the growth of bacteria and algae, remove harmful bacteria and algae in water, and improve water quality. This is helpful to the reduction of toxic substances such as ammonia nitrogen and nitrite in the water body, and plant polysaccharides can be used as a carbon source and energy source for microorganisms, promote the growth and reproduction of beneficial microorganisms, enhance the self-purification capacity of the water body, improve the dissolved oxygen content of the water body, improve the growth environment of the prawns, and reduce the occurrence of diseases.

Description

Technical Field

[0001] This invention relates to the field of aquaculture microbial technology, and in particular to a method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture. Background Technology

[0002] With the rapid development of my country's shrimp farming industry, shrimp farming holds an important position globally. However, with the expansion of farming scale, a series of problems have gradually emerged. The deterioration of water quality in the farming environment and the frequent occurrence of shrimp diseases have become problems that cannot be ignored, with excessive ammonia nitrogen in the farming water and the rampant spread of bacterial and vibrio diseases being two major issues. To address these problems, microecological preparations have gradually been widely used in shrimp farming. Microecological preparations mainly improve the farming environment and enhance the immunity of farmed organisms by introducing beneficial microorganisms, thereby reducing the occurrence of diseases. Currently, the existing microecological preparations on the market often have a relatively generalized effect and lack specialization. The production and processing techniques for microecological preparations are outdated and simplistic, with a lack of methods for post-production processing. This may lead to contamination of microecological preparations with other harmful bacteria during mixing with feed, compromising their safety and stability. Currently, in my country, the types of microbial strains suitable for shrimp-specific microecological preparations are relatively few. Furthermore, unsatisfactory microbial compatibility may affect the growth and reproduction conditions of the strains, hindering their effectiveness.

[0003] Compared to single microbial preparations, compound microecological preparations offer several advantages. They can synergistically improve the aquaculture environment by introducing multiple beneficial microorganisms. Furthermore, compound microecological preparations can enhance the immunity of farmed organisms and strengthen their resistance to disease. Therefore, this invention develops compound microecological preparations by screening and domesticating microbial strains suitable for shrimp farming, optimizing microbial compatibility, reducing ammonia nitrogen levels in water, effectively inhibiting Vibrio growth, improving shrimp immune function, and further enhancing seedling quality and seedling stability. Summary of the Invention

[0004] In view of this, the present invention proposes a method for preparing a compound microecological preparation for Litopenaeus vannamei farming, thereby solving the above problems.

[0005] The technical solution of this invention is achieved as follows: A method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture includes the following steps:

[0006] S1. Strains Selection: Select compound effective bacteria with probiotic effects and strong environmental adaptability, and activate them for 12-24 hours to ensure that the viable bacterial count reaches 1.52 × 10⁻⁶. 9 cfu / mL or higher;

[0007] S2. Enrichment culture: The activated compound effective bacteria are inoculated into the enrichment medium for enrichment culture. The inoculation amount is 1.2-1.8%. The culture process adopts the method of alternating between room temperature and low temperature culture for acclimatization. The alternation between room temperature and low temperature culture is carried out 2-4 times as a cycle, and the culture is carried out continuously for 3-5 cycles. The bacterial sediment is collected by centrifugation.

[0008] S3. Preparation of suspension: Take the above bacterial cell sediment and mix it with the protective solution, and stir it with ultrasound to make a suspension;

[0009] S4. Preparation: Take the suspension, antimicrobial peptide, and regulator in a mass-volume ratio of (25-55) mL:(1.5-5.5) g:(3.2-8.5) g and stir to mix. Mix well with a shaker and place in a refrigerator at -4 to 2℃ for 20-40 min. Then transfer to -20 to -10℃ and place for 50-70 min. Freeze-dry for 4-6 h to obtain the compound microecological preparation.

[0010] Furthermore, the effective bacteria in S1 are Clostridium butyricum solution, lactic acid bacteria solution, and Bacillus subtilis solution in a volume ratio of (4-6):(2-3):(1-3), and the viable count of Clostridium butyricum after activation is ≥1.8×10⁻⁶. 9 cfu / mL, viable count of lactic acid bacteria ≥2.2×10 9 CFU / mL, viable count of Bacillus subtilis ≥2.5 × 10⁻⁶ 9 cfu / mL.

[0011] Furthermore, the activation treatment of S1 involves inoculating the compound effective bacteria into the activation culture medium at an inoculation amount of 0.5-1.0%, controlling the activation temperature at 25-37℃, pH at 6-8, humidity at 80-90%, and light intensity at 1000-2000 lux.

[0012] Furthermore, the S2 culture temperature is 27-35℃ and the culture time is 6-10h, while the low temperature culture temperature is 13-15℃ and the culture time is 14-18h.

[0013] Furthermore, the enrichment medium for S2 is MS basal medium, glucose 5-10 g / L, sucrose 0.2-0.4 g / L, ammonium salt 1.0-1.5 mg / L, sulfate 1.2-2.2 mg / L, vitamin C 6-8 g / L, bile salt 0.05-0.1 mg / L, and rosehipic acid 0.03-0.08 mg / L.

[0014] Furthermore, the mass-to-volume ratio of the S3 bacterial cell deposits to the protective solution is 10-18:2 (g / mL).

[0015] Furthermore, the S3 protective solution comprises complexin, maltose, and glycerol in a mass-to-volume ratio of (3-10):(2-7):18 g / mL.

[0016] Furthermore, the ultrasonic stirring power of S3 is 0.5-1.2 kW / m³. 3 The ultrasonic frequency is 15-85kHz, and the ultrasonic treatment time is 0.5-1.5h.

[0017] Furthermore, the antimicrobial peptide of S4 is any one of cephalosporin, defensin, bee venom, bufotoxin, bacitracin, and brevicin S.

[0018] Furthermore, the regulator of S4 comprises the following raw materials in parts by weight: 12-25 parts of brown algae polysaccharide, 20-80 parts of asparagus polysaccharide, 10-15 parts of Phellodendron bark extract, 5-8 parts of perilla extract, and 2-5 parts of myrtle leaf.

[0019] Compared with the prior art, the beneficial effects of the present invention are:

[0020] This project develops a compound microecological preparation. Through screening and domestication, a multifunctional microecological preparation is developed. After the beneficial bacteria in the preparation enter the water, they exert their functions of oxidation, denitrification, phosphorus solubilization, sulfidation, and nitrogen fixation, rapidly decomposing organic matter such as farmed animal excrement, leftover feed, and plant and animal remains, effectively reducing the concentration of ammonia nitrogen and nitrite in the water. This helps maintain water cleanliness and stability, providing a favorable growth environment for shrimp and promoting the growth and reproduction of beneficial microorganisms in the water, such as single-celled algae. The photosynthesis of these microorganisms can increase the dissolved oxygen level in the water, providing more oxygen for the shrimp and contributing to their healthy growth. Through the improvement of water quality and the promotion of beneficial microorganisms, the compound microecological preparation can increase the yield of Pacific white shrimp. Simultaneously, the improved shrimp growth environment can also reduce the occurrence of diseases, further improving the quality of seedlings and the stability of seedling cultivation.

[0021] Compared to traditional farming methods, using compound microecological preparations in Litopenaeus vannamei farming can reduce the use of chemical drugs and decrease environmental pollution. This contributes to achieving environmentally friendly and sustainable farming practices, aligning with modern agricultural development trends. It offers multiple beneficial effects, including improving water quality, promoting the growth of beneficial microorganisms, increasing yields, and ensuring environmental sustainability. This provides a new, more environmentally friendly, and efficient farming method for Litopenaeus vannamei. Detailed Implementation

[0022] To better understand the technical content of this invention, specific embodiments are provided below to further illustrate the invention.

[0023] Unless otherwise specified, the experimental methods used in the embodiments of this invention are all conventional methods.

[0024] Unless otherwise specified, all materials and reagents used in the embodiments of this invention are commercially available.

[0025] Example 1

[0026] A method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture includes the following steps:

[0027] S1. Strains Selection: A compound of effective bacteria with probiotic properties and strong environmental adaptability was screened and inoculated into the activation medium at an inoculation rate of 0.5%. The activation temperature was controlled at 25℃, pH at 6, humidity at 80%, and light intensity at 1000 lux for 12 hours. The compound of effective bacteria consisted of Clostridium butyricum, Lactobacillus, and Bacillus subtilis in a volume ratio of 4:2:1. After activation, the viable count of the Clostridium butyricum was ≥1.8 × 10⁻⁶. 9 cfu / mL, viable count of lactic acid bacteria ≥2.2×10 9 CFU / mL, viable count of Bacillus subtilis ≥2.5 × 10⁻⁶ 9 cfu / mL;

[0028] S2. Enrichment Culture: The activated compound effective bacteria were inoculated into the enrichment medium at an inoculation rate of 1.2%. The culture process involved alternating between room temperature and low temperature culture for acclimatization. The room temperature culture temperature was 27℃ for 6 hours, and the low temperature culture temperature was 13℃ for 14 hours. This alternation of room temperature and low temperature culture was repeated twice as one cycle, and three consecutive cycles were cultured. The bacterial sediment was collected by centrifugation. The enrichment medium consisted of MS basal medium, glucose 5 g / L, sucrose 0.2 g / L, ammonium salt 1.0 mg / L, sulfate 1.2 mg / L, vitamin C 6 g / L, bile salt 0.05 mg / L, and rosehipic acid 0.03 mg / L.

[0029] S3. Preparation of suspension: Take the above bacterial cell sediment and protective solution and mix them ultrasonically at a mass-to-volume ratio of 10:2 (g / mL). The ultrasonic stirring power is 0.5 kW / m³. 3 The ultrasonic frequency was 15 kHz, the ultrasonic treatment time was 0.5 h, and a suspension was prepared. The protective solution included casein, maltose and glycerol with a mass-to-volume ratio of 3:2:18 g / mL.

[0030] S4. Preparation: Take the suspension, antimicrobial peptide, and regulator at a mass-volume ratio of 25mL:1.5g:3.2g and stir to mix. Mix well with a shaker. The antimicrobial peptide is cephalosporin. Place it in a refrigerator at -4℃ for 20 minutes, then transfer it to -20℃ for 50 minutes and freeze-dry for 4 hours to obtain a compound microecological preparation. The regulator includes the following raw materials in parts by weight: 12 parts of brown algae polysaccharide, 20 parts of asparagus polysaccharide, 10 parts of phellodendron bark extract, 5 parts of perilla extract, and 2 parts of myrtle leaf.

[0031] Example 2

[0032] A method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture includes the following steps:

[0033] S1. Strains Selection: A compound of effective bacteria with probiotic effects and strong environmental adaptability was screened and inoculated into the activation medium at an inoculation rate of 1.0%. The activation temperature was controlled at 37℃, pH at 8, humidity at 90%, and light intensity at 2000 lux for 24 hours. The compound of effective bacteria consisted of Clostridium butyricum, Lactobacillus, and Bacillus subtilis in a volume ratio of 6:3:3. After activation, the viable count of the Clostridium butyricum was ≥1.8 × 10⁻⁶. 9 cfu / mL, viable count of lactic acid bacteria ≥2.2×10 9 CFU / mL, viable count of Bacillus subtilis ≥2.5 × 10⁻⁶ 9 cfu / mL;

[0034] S2. Enrichment Culture: The activated compound effective bacteria were inoculated into the enrichment medium at an inoculation rate of 1.8%. The culture process involved alternating between room temperature and low temperature culture for acclimatization. The room temperature culture temperature was 35℃ for 10 hours, and the low temperature culture temperature was 15℃ for 18 hours. This alternation of room temperature and low temperature culture was repeated 4 times as one cycle, and 5 consecutive cycles were cultured. The bacterial sediment was collected by centrifugation. The enrichment medium consisted of MS basal medium, glucose 10 g / L, sucrose 0.4 g / L, ammonium salt 1.5 mg / L, sulfate 2.2 mg / L, vitamin C 8 g / L, bile salt 0.1 mg / L, and rosehipic acid 0.08 mg / L.

[0035] S3. Preparation of suspension: Take the above bacterial cell sediment and protective solution and mix them ultrasonically at a mass-to-volume ratio of 18:2 (g / mL). The ultrasonic stirring power is 1.2 kW / m³. 3 The ultrasonic frequency was 85kHz, the ultrasonic treatment time was 1.5h, and a suspension was prepared. The protective solution included casein, maltose and glycerol with a mass-to-volume ratio of 10:7:18 g / mL.

[0036] S4. Preparation: Take the suspension, antimicrobial peptide, and regulator at a mass-volume ratio of 55mL:5.5g:8.5g and stir to mix. Mix well with a shaker. The antimicrobial peptide is a defensin. Place it in a refrigerator at 2℃ for 40min, then transfer it to -10℃ for 70min and freeze-dry for 6h to obtain a compound microecological preparation. The regulator includes the following raw materials in parts by weight: 25 parts of brown algae polysaccharide, 80 parts of asparagus polysaccharide, 15 parts of Phellodendron bark extract, 8 parts of Perilla frutescens extract, and 5 parts of Myrtle leaf.

[0037] Example 3

[0038] A method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture includes the following steps:

[0039] S1. Strains Selection: A compound of effective bacteria with probiotic effects and strong environmental adaptability was screened and inoculated into the activation medium at an inoculation rate of 0.8%. The activation temperature was controlled at 32℃, pH 7, humidity 85%, and light intensity 1500 lux for 20 hours. The compound of effective bacteria consisted of Clostridium butyricum, Lactobacillus, and Bacillus subtilis in a volume ratio of 5:2.5:2. After activation, the viable count of Clostridium butyricum was ≥1.8 × 10⁻⁶. 9 cfu / mL, viable count of lactic acid bacteria ≥2.2×10 9 CFU / mL, viable count of Bacillus subtilis ≥2.5 × 10⁻⁶ 9 cfu / mL;

[0040] S2. Enrichment Culture: The activated compound effective bacteria were inoculated into the enrichment medium at an inoculation rate of 1.5%. The culture process involved alternating between room temperature and low temperature culture for acclimatization. The room temperature culture temperature was 32℃ for 8 hours, and the low temperature culture temperature was 14℃ for 16 hours. This alternation of room temperature and low temperature culture was repeated 3 times as one cycle, and 4 consecutive cycles were cultured. The bacterial sediment was collected by centrifugation. The enrichment medium was MS basal medium, glucose 8 g / L, sucrose 0.3 g / L, ammonium salt 1.2 mg / L, sulfate 1.7 mg / L, vitamin C 7 g / L, bile salt 0.07 mg / L, and rosehipic acid 0.05 mg / L.

[0041] S3. Preparation of suspension: Take the above bacterial cell sediment and protective solution and mix them ultrasonically at a mass-to-volume ratio of 14:2 (g / mL). The ultrasonic stirring power is 0.8 kW / m. 3 The ultrasonic frequency was 50 kHz, the ultrasonic treatment time was 1 h, and a suspension was prepared. The protective solution included complexin, maltose and glycerol with a mass-to-volume ratio of 8:5:18 g / mL.

[0042] S4. Preparation: Take the suspension, antimicrobial peptide, and regulator at a mass-volume ratio of 40mL:3.5g:5.5g and stir to mix. Mix well with a shaker. The antimicrobial peptide is bee venom. Place it in a refrigerator at -1℃ for 30 minutes, then transfer it to -15℃ for 60 minutes and freeze-dry for 5 hours to obtain a compound microecological preparation. The regulator includes the following raw materials in parts by weight: 18 parts of brown algae polysaccharide, 50 parts of asparagus polysaccharide, 12 parts of Phellodendron bark extract, 7 parts of Perilla frutescens extract, and 3 parts of Myrtle leaf.

[0043] Comparative Example 1

[0044] The difference between this comparative example and Example 3 is that, in the preparation process of the compound microecological preparation, the compound effective bacteria in step S1 were not activated.

[0045] Comparative Example 2

[0046] The difference between this comparative example and Example 3 is that in the preparation process of the compound microecological preparation, step S2 adopts a constant temperature enrichment culture, with a culture temperature of 32°C and a culture time of 24 hours.

[0047] Comparative Example 3

[0048] The difference between this comparative example and Example 3 is that, in the preparation process of the compound microecological preparation, antimicrobial peptides were not added for formulation in step S4.

[0049] Comparative Example 4

[0050] The difference between this comparative example and Example 3 is that, in the preparation process of the compound microecological preparation, no regulator was added for formulation in step S4.

[0051] I. Effect Test

[0052] The aquaculture area for Litopenaeus vannamei was divided into 7 experimental zones. The compound microecological preparations of Examples 1-3 and Comparative Examples 1-4 were added to the 7 experimental zones. The ammonia nitrogen, nitrite, and Vibrio content in the water before and after the addition were measured, and the removal efficiency and dissolved oxygen content after treatment were calculated.

[0053] Ammonia nitrogen removal rate (%) = 100% × (C o -C) / C o C o C represents the initial concentration of ammonia nitrogen in the water sample (mg / L), while C represents the remaining concentration of ammonia nitrogen in the water sample after treatment (mg / L).

[0054] Nitrite removal rate = (A0 - A) x ) / A0×100%; A0 represents the initial nitrite concentration; A x This represents the nitrite concentration after treatment;

[0055] Vibrio inhibition rate = (initial Vibrio concentration - post-treatment Vibrio concentration) / initial Vibrio concentration × 100%;

[0056] The dissolved oxygen was measured using the iodometric method.

[0057] The measurement results are shown in the table below:

[0058]

[0059] The above results show that the microecological preparation of the present invention can be rapidly dispersed and distributed in various parts of the water body, participate in the decomposition of fish uneaten food, feces and organic matter in the water, thereby accelerating the improvement of the water environment, helping to inhibit the reproduction and growth of harmful bacteria in the water, and effectively reducing the overall level of nitrite and ammonia nitrogen in the water by decomposing various organic substances, effectively inhibiting the growth of Vibrio, and creating a healthier growth environment for Litopenaeus vannamei.

[0060] Compared with Comparative Example 1, the activation of the compound effective bacteria activated the activity of the bacterial community. After the strains were activated, the resulting microbial community helped to form a benign cycle ecosystem, promoted the cycle and balance of ammonia and nitrogen in the water, and enhanced the self-purification capacity of the water.

[0061] Compared with Comparative Example 2, variable temperature culture can provide the optimal temperature conditions required for the growth of the strain at different temperature stages, thereby accelerating the growth rate and metabolic activity of the strain, simulating the temperature changes of the strain in the natural environment, thereby improving the strain's adaptability to environmental changes and enhancing its stress resistance and stability.

[0062] Compared with Comparative Example 3, the addition of antimicrobial peptides showed better water solubility, larger molecular weight, and broader antimicrobial activity. This effectively inhibited the growth of bacteria and algae, eliminating harmful bacteria and algae from the water to improve water quality. This helps reduce toxic substances such as ammonia nitrogen and nitrite in the water, promotes the reproduction and growth of food organisms such as diatoms and green algae, while inhibiting the reproduction of harmful algae, thus promoting the balance of the aquatic ecosystem, forming a healthy microbial community, and improving the self-purification capacity of the water body.

[0063] Compared with Comparative Example 4, the addition of regulators can remove pollutants such as heavy metal ions, organic matter, and ammonia nitrogen from water through adsorption and complexation mechanisms, improve water quality, and inhibit algae growth. This can effectively control the number of algae in the water and prevent problems such as algal blooms. The plant polysaccharides in the regulators can serve as carbon and energy sources for microorganisms, promote the growth and reproduction of beneficial microorganisms, enhance the self-purification capacity of the water, promote photosynthesis of plants in the water, increase the dissolved oxygen content of the water, and improve the ecological environment of the water.

[0064] II. Aquaculture Situation

[0065] The compound microecological preparations used in Example 3 and Comparative Examples 1-4 were employed in conventional aquaculture of Litopenaeus vannamei. The dosage of the microecological preparations was 3-5 ml / m³ per acre of pond. 3 Apply once every 5 to 7 days.

[0066] The yield and survival rate of the finished Litopenaeus vannamei shrimp were tested and statistically analyzed. The results are as follows:

[0067] Stocking density per acre (10,000 fish) Production (kg) Survival rate (%) Example 3 10 5122 95.33 Comparative Example 1 11 4976 88.42 Comparative Example 2 12 5021 89.75 Comparative Example 3 10 4806 83.64

[0068] Under the same conventional aquaculture model, different compound microecological preparations of the present invention were sprayed. The results show that the compound microecological preparation of Example 3 can promote the growth of beneficial microorganisms in the shrimp intestines, inhibit the growth and reproduction of harmful microorganisms, kill harmful bacteria, thereby adjusting the balance of beneficial microorganisms in the gastrointestinal tract, improving immunity and disease resistance, promoting shrimp growth, and increasing shrimp yield and survival rate.

[0069] 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 compound microecological preparation for Litopenaeus vannamei aquaculture, characterized in that: Includes the following steps: S1. Strains Selection: Select compound effective bacteria with probiotic effects and strong environmental adaptability, and activate them for 12-24 hours to ensure that the viable bacterial count reaches 1.52 × 10⁻⁶. 9 cfu / mL or higher; S2. Enrichment culture: The activated compound effective bacteria are inoculated into the enrichment medium for enrichment culture. The inoculation amount is 1.2-1.8%. The culture process is carried out by alternating between room temperature and low temperature culture. The alternation of room temperature and low temperature culture is carried out 2-4 times as one cycle, and the culture is carried out continuously for 3-5 cycles. The bacterial sediment is collected by centrifugation. S3. Preparation of suspension: Take the above bacterial cell sediment and mix it with the protective solution, and stir it with ultrasound to make a suspension; S4. Preparation: Take the suspension, antimicrobial peptide, and regulator at a mass-volume ratio of 25-55mL:1.5-5.5g:3.2-8.5g and stir to mix. Mix well with a shaker and place in a refrigerator at -4~2℃ for 20-40min. Then transfer to -20~-10℃ and place for 50-70min. Freeze-dry for 4-6h to obtain the compound microecological preparation. The effective bacteria in S1 consist of Clostridium butyricum solution, lactic acid bacteria solution, and Bacillus subtilis solution in a volume ratio of 4-6:2-3:1-3. After activation, the viable count of Clostridium butyricum is ≥1.8×10⁻⁶. 9 cfu / mL, viable count of lactic acid bacteria ≥2.2×10 9 CFU / mL, viable count of Bacillus subtilis ≥2.5 × 10⁻⁶ 9 cfu / mL; The S2 culture temperature is 27-35℃ and the culture time is 6-10h, while the low temperature culture temperature is 13-15℃ and the culture time is 14-18h. The enrichment medium in S2 is MS basal medium, glucose 5-10 g / L, sucrose 0.2-0.4 g / L, ammonium salt 1.0-1.5 mg / L, sulfate 1.2-2.2 mg / L, vitamin C 6-8 g / L, bile salt 0.05-0.1 mg / L, and rhodopsin 0.03-0.08 mg / L. The regulator in S4 includes the following raw materials in parts by weight: 12-25 parts of brown algae polysaccharide, 20-80 parts of asparagus polysaccharide, 10-15 parts of Phellodendron bark extract, 5-8 parts of Perilla frutescens extract, and 2-5 parts of Myrtle leaf.

2. The preparation method of the compound microecological preparation for Litopenaeus vannamei aquaculture as described in claim 1, characterized in that: The activation treatment in S1 involves inoculating the compound effective bacteria into the activation culture medium at an inoculation amount of 0.5-1.0%, controlling the activation temperature at 25-37℃, pH at 6-8, humidity at 80-90%, and light intensity at 1000-2000 lux.

3. The preparation method of the compound microecological preparation for Litopenaeus vannamei aquaculture as described in claim 1, characterized in that: The mass-to-volume ratio of bacterial sediment to protective solution in S3 is 10-18 g: 2 mL.

4. The preparation method of the compound microecological preparation for Litopenaeus vannamei aquaculture as described in claim 1, characterized in that: The protective solution in S3 comprises complexin, maltose, and glycerol in a mass-to-volume ratio of (3-10):(2-7):18, with units of g / mL.

5. The method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture as described in claim 1, characterized in that: The ultrasonic stirring power in S3 is 0.5-1.2 kW / m. 3 The ultrasonic frequency is 15-85kHz, and the ultrasonic treatment time is 0.5-1.5h.

6. The method for preparing a compound microecological preparation for Litopenaeus vannamei aquaculture as described in claim 1, characterized in that: The antimicrobial peptide in S4 is any one of cephalosporin, defensin, bee venom, bufotoxin, bacitracin, and brevicin.

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