A kind of aquaculture composite microbial agent and preparation method and preparation device thereof

Abstract

The present invention belongs to the technical field of aquaculture, and particularly relates to an aquaculture composite microbial inoculant and a preparation method and a preparation device thereof, comprising a fermentation kettle with a stirring component vertically built in, a microbial inoculant introduction component and a sampling component respectively arranged on the upper and lower parts of the fermentation kettle, and an output end of the microbial inoculant introduction component intersects with a rotating shaft stirring end of the stirring component in the fermentation kettle through a three-way pipe, a base is connected to the bottom of the fermentation kettle through a connecting column, and a sampling component is arranged between the bottom of the fermentation kettle and the base. The advantage of the present application is that uniform sampling of the preparation of the aquaculture composite microbial inoculant in the fermentation kettle is achieved by rotating a driving component, and the quantitative extraction of the sampled microbial inoculant can be achieved by combining the sampling component with the driving component at the same time; the gas used for supplying oxygen to the fermentation is linked with the delivery of the microbial liquid material, and the caliber of the liquid cavity is adjusted by the size of the gas intake volume, thereby compensating for the insufficient pressure supply to the liquid cavity.

Description

Technical Field

[0001] The invention belongs to the technical field of aquaculture, and in particular relates to an aquaculture composite microbial agent and a preparation method and a preparation device thereof. Background Art

[0002] At present, the aquaculture industry mainly uses physical, chemical, and biological-ecological methods to improve and repair the aquaculture water environment, and a series of water quality improver products have been derived from this. Chemical remediation is a method of eliminating or reducing pollutants in water through chemical reactions, but although chemical products act quickly, they lack long-term effectiveness, some products will cause secondary pollution, and some have high use costs.

[0003] Bioremediation is the use of specific organisms to eliminate or enrich environmental pollutants under certain conditions, thereby repairing and improving the polluted water environment. Using beneficial microorganisms to regulate the aquaculture water environment and purify water quality has attracted more and more attention. Various microorganisms have been gradually applied in water quality improvement and other aspects. These microorganisms can quickly degrade organic matter entering the aquaculture pond and effectively reduce ammonia nitrogen, sulfide, nitrite nitrogen, COD, etc. in the water, thereby effectively improving water quality and maintaining a good ecological environment in aquaculture waters.

[0004] At present, most of the microbial agents prepared by bioremediation methods on the market are single-bacteria fermentation products, or composite bacteria that are fermented separately and then mixed. When used in practice, multiple strains are prone to mutual incompatibility and cannot form a competitive advantage. The water purification effect is poor and it is impossible to stably maintain a good ecological environment in aquaculture waters. In addition, the fermentation process is the core step in the production and preparation of microbial agents, and its monitoring is related to product quality and output. Traditional fermentation monitoring methods rely on periodic manual sampling. However, this method has problems such as randomness of sampling positions, fixed sampling intervals, and insufficient sample representativeness, which makes it difficult to accurately reflect the overall microbial growth status and metabolic activity changes in the fermenter. In actual operation, the most difficult problem in the transmission of small batches of microbial agents is how to avoid uneven feeding and inefficiency caused by small bacterial dosage during pipeline transportation. When handling small batches of liquid microbial agents, the presence of gaps makes the flow of microbial agents in the pipeline unstable because the bacterial dosage is not enough to fill the entire transmission pipeline, making it difficult to ensure continuous and uniform supply, which directly affects the inoculation density and fermentation effect of the bacteria in the fermenter. Summary of the invention

[0005] The present invention provides a composite microbial agent for aquaculture and a preparation method and a preparation device thereof. Multiple strains in the composite microbial agent of the present invention can closely cooperate and mutually benefit from each other, which greatly improves the effect of water purification. The composite microbial agent can also be added to bait as a bait additive to promote the growth and development of aquaculture organisms, thus solving the above-mentioned problems.

[0006] The technical solution of the present invention is achieved in this way:

[0007] A composite microbial agent for aquaculture, comprising the following components by weight:

[0008] 15-800 parts of boronophilic Bacillus; 1-800 parts of Pichia pastoris; 1-800 parts of Bifidobacterium longum; 10-750 parts of yeast; 10-830 parts of photosynthetic bacteria; 10-850 parts of lactic acid bacteria; 8-800 parts of nitrifying bacteria; 8-850 parts of denitrifying bacteria; 5-700 parts of sulfur-producing Bacillus; 5-700 parts of volcanic bacillus; 5-600 parts of actinomycetes; 3-600 parts of halophilic bacteria; 3-800 parts of filamentous bacteria.

[0009] Based on the above technical scheme, Borophilic Bacillus: Gram-positive by Gram staining, rod-shaped cell morphology; round colonies, smooth and neat edges, milky white color, measured growth temperature range between 30℃-37℃, pH range of 6-11, salt concentration of 0-7%; in aquaculture, it can produce amylase, and can use organic matter produced by Pichia pastoris or yeast feed, as well as soluble starch in other organic matter such as feed, to provide fish with smaller molecular nutrients, promote fish absorption and growth. Pichia pastoris: This yeast is widely used in the field of biotechnology, such as as a host cell for expressing exogenous genes, used to produce various proteins and peptides, which is beneficial to aquaculture to increase nutrients and provide nutrients. Bifidobacterium longum and Pichia pastoris: These two strains can regulate the microecological balance in the intestines of fish, promote fish digestion and absorption, and improve feed utilization. At the same time, they can also produce some substances with antibacterial effects, which help to improve the disease resistance of fish. Lactic acid bacteria use nitrogen compounds obtained from photosynthetic bacteria and yeast as a matrix to produce lactic acid. Lactic acid has a strong bacteriostatic ability and can effectively inhibit the activity of harmful microorganisms and the rapid decomposition of organic matter. Lactic acid bacteria can make difficult-to-decompose organic matter such as lignin and cellulose easy to decompose, and eliminate the harmful effects caused by undecomposed organic matter. On the basis of organic matter fermentation and decomposition, it plays an important role. Under normal circumstances, if pathogenic bacteria increase, farmed fish and shellfish will become weak and grow slowly. Lactic acid bacteria can inhibit the activity of pathogenic bacteria and improve the health and activity of fish and shellfish. Yeast: Yeast is a single-cell fungus that can decompose organic matter in bait, such as protein, fat and sugar, and produce a large amount of nutrients, such as amino acids, vitamins and minerals. These nutrients can be absorbed and utilized by fish to promote the growth and development of fish. At the same time, yeast can also produce some substances with antibacterial effects, such as antibiotics and enzymes, which help to improve the disease resistance of fish and reduce the occurrence of diseases. Yeast is widely used in the fields of food, beverages, medicine and biotechnology. It can help digestion, regulate intestinal flora, and also has many functions such as beauty and skin care, assisting in regulating blood sugar and blood lipids, and improving immunity. Photosynthetic bacteria: This is a microorganism that uses light as energy and carbon dioxide or small molecular organic matter as a carbon source for growth. Photosynthetic bacteria is a type of bacteria that can photosynthesize. It can use light energy to convert carbon dioxide and water into organic matter and oxygen. In aquaculture water, photosynthetic bacteria can provide oxygen and nutrition for fish and maintain the ecological balance in the water. Photosynthetic bacteria can improve water quality, promote the growth of fish and shrimp, and increase production. At the same time, photosynthetic bacteria can also decompose harmful substances in water, such as ammonia nitrogen, nitrite and sulfide, to improve water quality and reduce toxicity to fish.

[0010] H2S+2H20+2C02 photosynthetic bacteria→2(HCHO)+H2SO4

[0011] Hydrogen sulfide + water + carbon dioxide photosynthetic bacteria → sugars + sulfuric acid

[0012] H2SO4+2NH3→(NH4)2SO4→Fertilizer

[0013] Sulfuric acid + ammonia → ammonium sulfate → fertilizer

[0014] Lactic acid bacteria: They can make full use of the probiotics in the formula and grow rapidly. They can attach to the epithelial cells of the intestinal villi in a short time, effectively form biofilms, and prevent the invasion of pathogens. They can also quickly secrete lactic acid and acetic acid, create an acidic environment below pH 4.8 in the intestinal cavity, and effectively inhibit the proliferation of pathogens. The metabolites are rich in vitamin B group, amino acids, etc., which are the most basic and important elements for improving immunity. Nitrifying bacteria and denitrifying bacteria: Nitrifying bacteria can convert ammonia nitrogen into nitrite. Nitrifying bacteria are an autotrophic species. Denitrifying bacteria can reduce nitrite to nitrogen gas and discharge it into the atmosphere. These two bacteria can reduce ammonia nitrogen and nitrite in water bodies, can carry out nitrogen circulation in water bodies, reduce the concentration of ammonia nitrogen in water bodies, and reduce toxicity to fish. In addition, nitrifying bacteria can also produce some substances with antibacterial effects, such as nitrification enzymes and nitrification inhibitors, which help improve the disease resistance of fish. Sulfur-producing Bacillus: This strain is related to the conversion and utilization of sulfur. 1. Acid resistance: The metabolism is adapted to acidic environments, and it can survive and reproduce under low pH conditions. 2. Sugar metabolism: It has a variety of sugar metabolism pathways. It can use a variety of different carbon sources, such as glucose, fructose, lactose, etc., and convert these carbon sources into energy and organic acids through the glycolysis pathway. 3. Lactic acid metabolism: In an acidic environment, lactic acid fermentation may be carried out to convert carbon sources such as glucose into lactic acid. Make it easy to decompose difficult-to-decompose organic matter 4. Amino acid metabolism: It can use amino acids as carbon and energy. It has an amino acid degradation pathway to produce organic acids and ammonia by decomposing amino acids. 5. Spore formation: Like other Bacillus, it can form spores that tolerate extreme conditions to survive in unsuitable environments. This helps it survive during processing. Volcanic Bacillus: A Gram-positive rod of the class Bacillus. Because it is found in extreme environments such as volcanoes, this strain has special physiological functions and adaptability such as heat resistance, which can produce antibacterial substances, inhibit the reproduction of harmful microorganisms, degrade nutrients, and improve the ecological environment. It can produce bacterial protease. It can be used to reduce nitrates and hydrolyze starch. It can decompose indigestible substances such as cellulose and protein in bait and improve the utilization rate of bait. At the same time, Bacillus can also produce some substances with antibacterial effects, such as antibiotics and enzymes, which help to improve the disease resistance of fish. In addition, Bacillus can also promote the growth of beneficial bacteria in the intestines of fish and maintain the balance of intestinal microecology. Actinomycetes: Actinomycetes are Gram-positive bacteria that can decompose organic substances in water bodies, such as proteins, fats and sugars, and produce a large amount of nutrients, such as amino acids, vitamins and minerals. These nutrients can be absorbed and utilized by fish and promote the growth and development of fish. At the same time, actinomycetes can produce various antibiotics, such as antibiotics and enzymes, which can directly inhibit harmful pathogens, help improve the disease resistance of fish, and create a living environment for the proliferation of other beneficial microorganisms.Since actinomycetes and photosynthetic bacteria can coexist, the antibacterial effect of actinomycetes and photosynthetic bacteria when mixed with actinomycetes alone is multiplied. Halophiles: Halophiles are bacteria that can survive in high-salinity environments. Halophiles are of certain significance to special species of fish such as farmed marine fish. They can decompose organic matter in bait in a high-salinity environment and produce a large amount of nutrients for fish to absorb and utilize. At the same time, halophiles can also maintain the ecological balance in the water body and reduce the occurrence of diseases. Filamentous fungi: Filamentous fungi are a type of multicellular fungi that can decompose organic matter in the water body, such as proteins, fats and sugars, and produce a large amount of nutrients, such as amino acids, vitamins and minerals. These nutrients can be absorbed and utilized by fish to promote their growth and development. At the same time, filamentous fungi can also produce some substances with antibacterial effects, such as antibiotics and enzymes, which help improve the disease resistance of fish.

[0015] A method for preparing a composite microbial agent for aquaculture, comprising the following steps:

[0016] S2. The culture medium for culturing the bacterial flora includes RS lipid, sodium oxide, Sabouraud dextrose agar (SDA) medium, potato dextrose agar, yeast extract, Tween 80 corn agar, broth, malt extract, vitamin C, vitamin D, lactose, defatted protein and TGY agar, and is sterilized and inactivated at a temperature of 70-80° C. for 5-40 minutes to prepare a composite culture medium for culturing the composite mother strain;

[0017] S3, adding the composite mother strain in step S1 to the composite culture medium that has been sterilized and inactivated in step S2, and incubating at a temperature of 10-60° C. for natural fermentation;

[0018] S4. The composite microbial agent obtained by cultivation in step S3 is sealed and stored in an environment of -45-0°C.

[0019] Based on the above technical solutions, the culture medium of the bacterial community and its fermentation products (the products of bacterial community fermentation include amino acids, organic acids, polysaccharides, inorganic salts, etc.) and various trace element fermentation products have the following functions: probiotics effectively decompose metabolites (organic matter), proteins, fibers, hemicelluloses, and α-amylases produced in aquaculture. It enables aquaculture water to effectively absorb feed and increase production capacity. Symbiotic and mutually beneficial effect: Many compound probiotic products on the market are produced through separate culture and fermentation models, that is, freeze-dried and made into a single bacterial powder; or a variety of bacterial powders are mixed together to make a commercially available "compound probiotic"; this model is low-cost, and the probiotics produced not only have a low survival rate, but also fail to retain the beneficial metabolites produced during the microbial fermentation process. The present invention selects highly active excellent strains with the strongest specific functional performance, and co-cultivates several carefully selected composite strains, which coexist and complement each other to form the principle of symbiotic and coexisting beneficial bacteria, that is, "the more strains, the better the survival rate"; after fermentation, all the bacterial liquid and metabolites are freeze-dried together, which not only retains the efficacy of the bacteria, but also retains the extremely beneficial metabolites, so that all bacteria can play the greatest benefit to the human body. The composite probiotics produced in this mode can support each other and are less likely to be wiped out in one fell swoop, and the approval protection effect of the composite bacteria is much better than that of a single strain. Wide temperature adaptability: Insulation cultivation is carried out in a temperature range of 10-60°C, which shows that this preparation method has good temperature adaptability. This means that microorganisms can be effectively cultivated in a warmer or cooler environment, which increases the flexibility and practicality of the preparation process. Natural fermentation process: The natural fermentation method can better simulate the growth conditions of microorganisms in the natural environment, which helps to maintain the activity and stability of microorganisms. At the same time, the natural fermentation process does not require the addition of additional chemical reagents or catalysts, which is more environmentally friendly and safe. Flexible storage conditions: The composite microbial inoculant can be stored in a sealed state at -45-0°C. This wide temperature range makes the storage and transportation of the inoculant more convenient. Whether it is stored for a long time under frozen conditions or for a short time at room temperature or slightly lower temperatures, the activity and effect of the inoculant can be guaranteed. Efficient and easy to operate: The entire preparation process is relatively simple and does not require complex equipment or technology, making this method easy to apply and promote in actual production. At the same time, by optimizing the cultivation conditions and storage methods, the efficiency and stability of the composite microbial inoculant can be ensured.

[0020] Application of composite microbial agents in aquaculture for water purification.

[0021] Application of composite microbial agents in aquaculture to promote the growth of aquatic organisms.

[0022] A device for preparing a composite microbial agent for aquaculture, comprising:

[0023] A fermentation kettle with a stirring assembly built in vertically, a bacterial agent introduction assembly and a sampling assembly are respectively arranged on the upper and lower parts of the fermentation kettle, and the output end of the bacterial agent introduction assembly is connected with the stirring end of the rotating shaft of the stirring assembly through a three-way pipe in the fermentation kettle;

[0024] The bottom of the fermentation kettle is connected to the base through a connecting column, and the sampling component is arranged between the bottom of the fermentation kettle and the base. It is specifically a rotating drive component driven by a first motor, and an interveneable manual drive component is added to the rotating drive component, thereby realizing free switching between automatic sampling and manual sampling.

[0025] Preferably, a plurality of outlets are equidistantly provided at the bottom of the fermentation kettle along its circumference, and the sampling assembly is composed of a number of cylinders equal to the number of the outlets, and the cylinders are respectively arranged at each outlet; a first opening and closing member and a second opening and closing member of the same structure are respectively provided at the upper and lower ends of the cylinder. Taking the first opening and closing member as an example, it is composed of an upper opening and closing plate and a lower opening and closing plate hinged in a cross shape, a reset spring is provided at the hinge, and a trigger plate extends outward from any opening and closing plate, and the two opening and closing plates seal the end face of the cylinder in a closed state; a shell is also provided between the bottom of the fermentation kettle and the base, and the bottom of the cylinder is a sampling port, and the shell is used to isolate each sampling port from the rotating drive assembly.

[0026] Based on the above technical solutions, the sampling component and the rotation drive component work together to accurately control the sampling volume, ensure that the amount of bacterial agent sample exported each time is consistent, and provide a reliable quantitative basis for experimental analysis; the use of the first motor to drive the rotation drive component can accurately control the rotation speed and sampling position, which is suitable for large-scale automated production and improves sampling efficiency and accuracy. The motor-driven automated operation reduces the interference of human factors and ensures the stability and repeatability of the sampling process; in situations where flexible operation or emergency situations are required, the manual drive component provides a convenient solution, so that the sampling operation can be successfully completed even when the motor drive conditions are not available or temporary adjustments are required.

[0027] Preferably, the rotation drive assembly includes an inner gear ring which is connected as an integral body and has an upper and lower double layer, and the outer circumferential surfaces of the two inner gear rings are respectively connected to a first lever and a second lever, the two levers are arranged in a staggered manner, and the first lever and the second lever are respectively in the same horizontal plane as the trigger plates of the first opening and closing member and the second opening and closing member; the rotation drive assembly also includes a second rotating gear connected to the output end of the first motor, and a plurality of first rotating gears are circumferentially meshed with the second rotating gear and the inner gear ring, and the thickness of the first rotating gear is greater than the thickness of the inner gear ring.

[0028] Preferably, one end of the manual drive member is rotatably connected to the base, which includes an integrated worm and handwheel, both of which are rotatably connected to the translation block, and the worm corresponds to any first rotating gear. Two elastic limiting protrusions are provided on the base, which correspond to the circular grooves at the bottom of the translation block, and are used to fix the two states of engagement and disengagement between the worm and the first rotating gear.

[0029] Preferably, the bacterial agent introduction component includes an introduction shell and a guide tube rotatably arranged in the introduction shell, an air inlet is provided on the side of the introduction shell, a liquid chamber and a gas chamber are arranged side by side inside the guide tube along its length direction, the axial guide of the liquid chamber is placed at the center, and the gas chamber faces one side of the three-way pipe and is in a conducting state; a conducting port is provided at the guide tube corresponding to the air inlet, and an axial sliding plate is provided at the conducting port to control the caliber and the air intake amount by movement; both ends of the guide tube are arranged as caliber fixed tubes, and the middle is arranged as a radial telescopic tube with variable diameter between the liquid chamber and the gas chamber, an arc spring for automatic reset is built in the radial telescopic tube, a filter is installed between the end of the radial telescopic tube and the caliber fixed tube, pressure sensors are installed in the front and rear port caliber fixed tubes, and the radial telescopic tube adjusts the caliber of the liquid chamber by changing the air intake amount of the conducting port, thereby compensating for the insufficient pressure supply of the liquid chamber.

[0030] Based on the above technical solutions, more accurate and efficient oxygen supply control can be achieved by linking the gas and liquid material delivery systems. The diameter or pressure of the liquid cavity can be indirectly controlled by dynamically adjusting the gas intake volume, thereby optimizing the dispersion and oxygen mixing state of the bacterial liquid in the fermentation tank.

[0031] Preferably, a second motor is provided on the outside of the inlet shell, and its output end is meshed with a ring gear through a gear that passes through the guide tube at one end of the inlet shell, and its end is rotatably connected to the three-way tube; opposite external threads are provided on both sides of the guide tube, and the guide tube is respectively threaded to two axial sliding plates, and a concave-convex structure is provided between the axial sliding plates and the inlet shell to prevent synchronous turnover; deformation capsules are provided between the two ends of the radial telescopic tube and each caliber fixed tube, and after the radial telescopic tube is reduced in diameter, the deformation capsule is pressed inward to deform, thereby blocking the excess area of ​​each filter screen, while ensuring the sealing effect at the joint surface of the radial telescopic tube and the caliber fixed tube.

[0032] In summary, this application includes the following beneficial technical effects:

[0033] 1. The composite microbial agent of the present invention can improve the stench phenomenon at the bottom of the aquaculture pond and reduce the probability of disease in aquaculture organisms. The composite microbial agent can utilize nitrogen (NHD), nitrite (NOQ) and hydrogen sulfide produced by the decomposition of sludge at the bottom of the pond under anaerobic conditions to achieve the purpose of proliferation and detoxification, and compete with oxygen-consuming heterotrophic microorganisms for nutrient organic matter, which can reduce the biological oxygen demand at the bottom of the pond, thicken the oxidation layer, and improve the oxidation potential, thereby achieving the purpose of purifying the bottom of the pond and water quality; the composite microbial agent of the present invention can also remove toxic substances in water, purify water quality, stabilize water quality, have strong cation exchange capacity, can effectively remove nitrogen, nitrite and hydrogen sulfide, have high stability, and long-term application will not cause accumulation and pollution of organic matter in the pond, solve the acidification of the bottom of the pond, improve the deterioration of the bottom of the pond, activate the dominant bacteria, and regenerate the pond environment.

[0034] 2. The composite microbial agent of the present invention can also remove toxic substances in water, purify water quality, stabilize water quality, has strong cation exchange capacity, can effectively remove nitrogen, nitrite and hydrogen sulfide, has high stability, and long-term application will not cause accumulation and pollution of organic matter in the pool, solve the acidification of the pool bottom, improve the deterioration of the pool bottom, activate the dominant bacterial community, and regenerate the pool environment; the composite microbial agent of the present invention can survive in polluted waters, compete with other pathogens for nutrient sources, use the advantages of the bacterial community to grab nutrients, so that the pathogens are inhibited due to nutrient deficiency, thereby achieving the effect of preventing the occurrence of diseases, and can increase the nutrients of animal plankton, and use bait to reduce the investment cost of artificial breeding.

[0035] 3. The present invention realizes uniform sampling of the preparation of aquaculture composite microbial inoculants in a fermentation kettle through a rotating drive component. The simultaneous use of the rotating drive component in combination with the sampling component can realize quantitative extraction of the sampled inoculants. The power conversion of the rotating drive component can be realized through the first motor drive and the manual drive component.

[0036] 4. The present invention links the gas used to supply oxygen for fermentation with the delivery of the bacterial agent liquid material, adjusts the caliber of the liquid cavity by the size of the gas intake volume, and thus compensates for the insufficient pressure supply of the liquid cavity. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative labor.

[0038] Figure 1 It is the change of inorganic nitrogen in the water of the aquaculture pond after applying the composite microbial agent;

[0039] Figure 2 It is the COD in the aquaculture pond water after applying the composite microbial agent. Mn changes;

[0040] Figure 3 It is the change of total heterotrophic bacteria and Vibrio in the water of the aquaculture pond after applying the compound microbial agent;

[0041] Figure 4 It is a schematic diagram of the structure of the preparation device in the present invention;

[0042] Figure 5 The present invention Figure 4 Exploded schematic diagram of the structure;

[0043] Figure 6 The present invention Figure 4 A schematic cross-sectional view of the bottom structure with the shell;

[0044] Figure 7 The present invention Figure 4 A schematic cross-sectional view of the bottom structure without the shell;

[0045] Figure 8 The present invention Figure 7 A schematic cross-sectional view of the structure from above;

[0046] Fig. 9 Schematic diagram of the explosion of the rotary drive assembly in the preparation device of the present invention Figure 1 ;

[0047] Fig.10 Schematic diagram of the explosion of the rotary drive assembly in the preparation device of the present invention Figure 2 ;

[0048] Fig.11 It is a cross-sectional schematic diagram of the sampling assembly in the preparation device of the present invention in different states;

[0049] Fig.12 The present invention Figure 4 A schematic cross-sectional view of the bottom structure;

[0050] Fig.13 It is a schematic diagram of the structure of the bacterial agent introduction component after opening and closing in the preparation device of the present invention;

[0051] Fig.14 The present invention Fig.13 Schematic cross-section of the structure;

[0052] Fig.15 The present invention Fig.13 Exploded schematic diagram of the structure;

[0053] Fig.16 The present invention Fig.15 Schematic diagram of the structure after removing the import shell;

[0054] Fig.17 The present invention Fig.16 Cross-section diagram of the structure Figure 1 ;

[0055] Fig.18 The present invention Fig.16 Cross-section diagram of the structure Figure 2 ;

[0056] Fig.19 The present invention Fig.16 Cross-section diagram of the structure Figure 3 ;

[0057] Fig. 20 It is an exploded schematic diagram of the radial telescopic tube and the filter screen in the present invention;

[0058] Fig.21 The cross-sectional view of the radial telescopic tube structure of the present invention is shown in FIG. Figure 1 ;

[0059] Fig. 22 The cross-sectional view of the radial telescopic tube structure of the present invention is shown in FIG. Figure 2 .

[0060] Description of reference numerals:

[0061] 1. Fermentation kettle; 2. Bacteria introduction component; 3. Stirring component; 4. Sampling component; 5. Manual drive component; 6. Rotation drive component; 10. Shell; 11. Observation window; 12. Three-way pipe; 13. First motor; 101. Connecting column; 102. Base; 103. Limiting convex point; 201. Introduction shell; 202. Guide pipe; 203. Second motor; 204. Axial sliding plate; 205. Filter; 206. Radial telescopic tube; 207. Deformation capsule; 2011. Air inlet; 2012. Limiting convexity; 2021. Liquid cavity; 2022. Gas cavity; 2023. Ring gear; 2024. External Thread; 2025, conducting port; 2026, positioning point; 2041, limiting through groove; 2061, L-shaped arc crimping plate; 2062, first arc plate; 2063, second arc plate; 2064, arc spring; 2065, annular deformation membrane; 401, cylinder; 402, first opening and closing member; 403, second opening and closing member; 4021, upper opening and closing plate; 4022, trigger plate; 4023, lower opening and closing plate; 501, worm; 502, translation block; 503, hand wheel; 601, inner gear ring; 602, first rotating gear; 603, second rotating gear; 604, first shifting rod; 605, second shifting rod. DETAILED DESCRIPTION

[0062] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0063] The embodiments of the present application disclose an aquaculture composite microbial agent and a preparation method and a preparation device thereof.

[0064] Example 1

[0065] A composite microbial agent for aquaculture, comprising the following components in parts by weight:

[0066] 15-800 parts of boronophilic Bacillus; 1-800 parts of Pichia pastoris; 1-800 parts of Bifidobacterium longum; 10-750 parts of yeast; 10-830 parts of photosynthetic bacteria; 10-850 parts of lactic acid bacteria; 8-800 parts of nitrifying bacteria; 8-850 parts of denitrifying bacteria; 5-700 parts of sulfur-producing Bacillus; 5-700 parts of volcanic bacillus; 5-600 parts of actinomycetes; 3-600 parts of halophilic bacteria; 3-800 parts of filamentous bacteria.

[0067] The concentration of nitrifying bacteria and denitrifying bacteria is high, which can more effectively remove ammonia nitrogen and nitrite in water and improve the water purification effect. The rich bacterial species diversity of the composite microbial agent contains many different types of microorganisms, including yeast, lactic acid bacteria, photosynthetic bacteria, Bacillus, actinomycetes, etc. These species work synergistically with each other to better play their respective functions and improve the water purification effect. Bifidobacterium longum and Pichia pastoris are added: these two probiotics have the effect of promoting intestinal health, can improve the nutritional value of bait, and enhance the immunity and disease resistance of fish. In addition to the role of water purification and bait, the composite microbial agent in the present invention may also have a certain protective effect on the environment. The photosynthetic bacteria can produce oxygen through photosynthesis, which helps to maintain the oxygen content in the water body; while Bacillus and actinomycetes can decompose organic matter and reduce the concentration of pollutants in the water body.

[0068] Example 2

[0069] A method for preparing a composite microbial agent for aquaculture, comprising the following steps:

[0070] S1. Mixing and stirring the above-mentioned multiple strains under sterile conditions to obtain a composite mother strain;

[0071] S2. The culture medium for culturing the bacterial flora includes RS lipid, sodium oxide, Sabouraud dextrose agar (SDA) medium, potato dextrose agar, yeast extract, Tween 80 corn agar, broth, malt extract, vitamin C, vitamin D, lactose, defatted protein and TGY agar, and is sterilized and inactivated at a temperature of 70-80° C. for 5-40 minutes to prepare a composite culture medium for culturing the composite mother strain;

[0072] S3, adding the composite mother strain in step S1 to the composite culture medium that has been sterilized and inactivated in step S2, and incubating at a temperature of 10-60° C. for natural fermentation;

[0073] S4. The composite microbial agent obtained by cultivation in step S3 is sealed and stored in an environment of -45-0°C.

[0074] Wherein, in step S1, according to the function and characteristics of each strain, and different metabolic pathways, the symbiotic and mutually beneficial effect is achieved. Then the selected composite mother strains are mixed and stirred to ensure that they are fully in contact and form a uniform mixture. The high temperature inactivation treatment in step S2 can kill the pathogenic microorganisms and other harmful organisms therein. The liquid culture medium in step S3 can provide the required nutrients and growth factors to promote the growth and reproduction of the flora. During the fermentation process, the flora will interact and produce beneficial metabolites. Insulation cultivation can be achieved in a constant temperature incubator or other appropriate equipment. In step S4, sealed storage can be used to prevent it from being affected by the external environment. This can be achieved by transferring the flora to a low temperature storage container and storing it in a sealed state.

[0075] Example 3

[0076] Select a culture pond, put grouper in a plurality of culture ponds for feeding, and detect the effect of the composite microbial agent in the present invention in purifying water quality and promoting the growth of grouper. The culture pond is divided into a control group and a test group. Add the composite microbial agent at the entrance of each culture pond and biofilter of the test group, 2 times a day, 1L each time, for a total of 19 days, and regularly detect the content indicators such as nitrite nitrogen, ammonia nitrogen, and organic matter in the water of the test group and the control group. On the 20th day, the test group stopped adding the composite microbial agent, and the time of ultraviolet light irradiation was reduced from 24h / day to 8h / day. From the 46th day, the daily feeding amount of the test group and the control group was increased from 15kg to 17.5kg, and the control group mixed the bait according to 10kg bait per 300mL. Wherein, the test of the culture pond was selected to be carried out in Workshop No. 7 of Yellow Sea Aquatic Products Co., Ltd., Haiyang City, Shandong Province.

[0077] from Figure 1It can be seen that during the entire test, the concentrations of ammonia nitrogen and nitrite in the test group were lower than those in the control group, with the concentration of ammonia nitrogen in the test group lower than 0.5 mg / L and the concentration of nitrite lower than 0.1 mg / L. After the bait was added, the concentrations of ammonia nitrogen and nitrite increased slightly, but were still within the safe range, and the concentrations of ammonia nitrogen and nitrite in the test group were still lower than those in the control group.

[0078] from Figure 2 It can be seen that during the entire experiment, the organic matter content of the experimental group was always lower than that of the control group. Moreover, although the organic matter content of the experimental group fluctuated, the fluctuation range of the organic matter content of the experimental group was smaller than that of the control group.

[0079] from Figure 3 It can be seen that during the entire period of feeding, the total heterotrophic bacteria in the experimental group increased, but the number of Vibrio decreased. During the period of stopping feeding, the total heterotrophic Vibrio number in the experimental group remained constant and had no significant difference from the control group. It can be seen that the experimental group was not affected by the shortening of UV light irradiation time. In addition, after the addition of bait, the total heterotrophic bacteria did not change significantly, and the number of Vibrio in the experimental group was lower than that in the control group.

[0080] After adding the compound microbial agent, the foam in the water was significantly reduced in the experimental group, and the activity and food intake of the grouper increased.

[0081] Table 1 is the growth and weight record of grouper

[0082] date 1 day 15 days 25 days Experimental Group 256.7g 391.2g 400.6g Control group 258.3g 373.0g 378.0g

[0083] As can be seen from Table 1, the growth rate of grouper in the experimental group was 56.1%, and the growth rate of grouper in the control group was 46.3%. It can be seen that the compound microbial agent can promote the growth and development of grouper after being added to the bait.

[0084] Select aquaculture ponds whose water quality does not meet the requirements for aquaculture water, add compound microbial agents to the aquaculture ponds every day, and test the water quality indicators after a period of time.

[0085] Table 2 Various indicators of water quality in the culture pond before and after adding composite microbial agents

[0086]

[0087]

[0088] It can be seen that after adding the compound microbial agent, the water quality in the breeding pond has been improved, indicating that the compound microbial agent has the effect of improving water quality.

[0089] Example 4

[0090] Based on the above embodiments, combined with Figures 4 to 12 This example also discloses a device for preparing a composite microbial agent for aquaculture, including a fermentation kettle 1 with a stirring component 3 vertically built in, a sampling component 4 is provided below the fermentation kettle 1, the bottom of the fermentation kettle 1 is connected to a base 102 through a connecting column 101, and the sampling component 4 is arranged between the bottom of the fermentation kettle 1 and the base 102, which is specifically a rotating drive component 6 driven by a first motor 13, and an intrusive manual drive component 5 is added to the rotating drive component 6.

[0091] A plurality of outlets are equidistantly provided at the bottom of the fermentation kettle 1 along its circumference. The sampling assembly 4 is composed of a number of cylinders 401 equal to the number of the outlets, and the cylinders 401 are respectively arranged at each outlet. In this structure, a first opening and closing member 402 and a second opening and closing member 403 with the same structure are respectively provided at the upper and lower ends of the cylinder 401. Taking the first opening and closing member 402 as an example, it is composed of an upper opening and closing plate 4021 and a lower opening and closing plate 4023 hinged in a cross shape, and a reset spring is provided at the hinge. A trigger plate 4022 is extended outwardly from any opening and closing plate, and the two opening and closing plates seal the end face of the cylinder 401 in a closed state.

[0092] A shell 10 is also provided between the bottom of the fermentation kettle 1 and the base 102. The bottom of the cylinder 401 is a sampling port. The shell 10 is used to isolate each sampling port from the rotating drive component 6. The inner interlayer of the fermentation kettle 1 is provided with a conventional temperature control structure for bacterial agent fermentation. In this structure, an observation window 11 is also provided on the fermentation kettle 1. A sampling cylinder can be placed laterally at the bottom of the cylinder 401 to achieve sampling of the bacterial agent. A corresponding pressure sensor can be provided at the corresponding position of the sampling cylinder on the base 102. When it is detected that the sampling cylinder is placed in the designated area, the rotating drive component 6 is controlled to rotate to the corresponding position to perform automatic liquid discharge sampling of the bacterial agent.

[0093] The rotation drive assembly 6 includes an inner gear ring 601 that is connected as an integral body and has an upper and lower double layer. The outer circumferential surfaces of the two inner gear rings 601 are respectively connected with a first lever 604 and a second lever 605. The two levers are arranged in a staggered manner, and the second lever 605 and the first lever 604 are respectively in the same horizontal plane with the trigger plates 4022 of the first opening and closing member 402 and the second opening and closing member 403. The rotation drive assembly 6 also includes a second rotating gear 603 connected to the output end of the first motor 13. A plurality of first rotating gears 602 are circumferentially meshed between the second rotating gear 603 and the inner gear ring 601, and the thickness of the first rotating gear 602 is greater than that of the inner gear ring 601. In this structure, the first lever 604 and the second lever 605 both have an electric telescopic function. When the lever is extended, it can act on the opening and closing member to realize opening and closing. Anyway, when the lever is not extended, the rotation drive assembly 6 rotates and does not act on the opening and closing member to perform opening and closing operations.

[0094] One end of the manual drive member 5 is rotatably connected to the base 102, which includes an integrated worm 501 and a hand wheel 503, both of which are rotatably connected to the translation block 502, and the worm 501 corresponds to any first rotating gear 602. Two elastic limiting protrusions 103 are provided on the base 102, which correspond to the circular grooves at the bottom of the translation block 502, and are used to fix the two states of engagement and disengagement between the worm 501 and the first rotating gear 602. In this structure, after the worm 501 is engaged with the first rotating gear 602, the first motor 13 is turned off to rotate the second rotating gear 603. At this time, the first rotating gear 602 acts as a driving gear to control the rotation of the inner gear ring 601.

[0095] The specific implementation process is: when automatic sampling is required, the first motor 13 is triggered by placing the sampling tube on the base 102, and the second rotating gear 603 rotates to drive the first rotating gear 602 and the inner gear ring 601 to rotate, so that the second lever 605 and the first lever 604 act on the first opening and closing member 402 and the second opening and closing member 403 respectively; the first opening and closing member 402 is arranged at the top, and the second opening and closing member 403 is arranged at the bottom. First, the second lever 605 at the top acts on the trigger plate 4022 to open the first opening and closing member 402, so that the bacterial agent enters the cylinder 401 and is filled; then the first lever 604 acts on the trigger plate 4022 to open the second opening and closing member 403. At this time, the first opening and closing member 402 is closed under the action of the reset spring, thereby realizing the sampling and export of the bacterial agent in the cylinder 401.

[0096] Example 5

[0097] Based on the above embodiments, combined with Figures 13 to 22 This example also discloses a preparation device for a composite microbial agent for aquaculture, which also includes an agent introduction component 2 provided above the fermentation kettle 1, the output end of the agent introduction component 2 intersects with the rotating shaft stirring end of the stirring component 3 through the three-way pipe 12 in the fermentation kettle 1, and the agent introduction component 2 in this structure includes an introduction shell 201 and a guide tube 202 rotatably arranged in the introduction shell 201, an air inlet 2011 is opened on the side of the introduction shell 201, and a liquid cavity 2021 and a gas cavity 2022 are arranged in parallel along the length direction of the guide tube 202, the liquid cavity 2021 is axially connected and placed at the center position, and the gas cavity 2022 is in a conductive state facing one side of the three-way pipe 12, and a conduction port 2025 is opened at the guide tube 202 corresponding to the air inlet 2011, and an axial sliding plate 204 is provided at the conduction port 2025 to control the caliber and the air intake volume by movement;

[0098] The two ends of the flow guide tube 202 are set as fixed-diameter tubes, and the middle is set as a radial telescopic tube 206 with a variable diameter between the liquid chamber 2021 and the gas chamber 2022. The radial telescopic tube 206 is built with an arc spring 2064 for automatic reset. A filter 205 is installed between the end of the radial telescopic tube 206 and the fixed-diameter tube. Pressure sensors are installed in the front and rear fixed-diameter tubes. The radial telescopic tube 206 adjusts the diameter of the liquid chamber 2021 by changing the size of the air intake of the guide port 2025, thereby compensating for the insufficient pressure supply of the liquid chamber 2021. In this structure, the front and rear pressure sensors are used to detect whether the filter screen 205 has insufficient pressure supply; the radial telescopic tube 206 is composed of a first arc plate 2062 and a second arc plate 2063 alternately arranged in the circumferential direction, the first arc plate 2062 is circumferentially slidably inserted in the second arc plate 2063, and a plurality of arc springs 2064 are arranged between the two, and a plurality of first arc plates 2062 and second arc plates 2063 form an annular surface, and an annular deformation membrane 2065 is sleeved on the inner side, and the annular deformation membrane 2065 is used to prevent the bacterial agent from contacting the joint surface of the arc plate.

[0099] A second motor 203 is provided on the outside of the introduction shell 201, and its output end is meshed with the ring gear 2023 that passes through one end of the introduction shell 201 through a gear, and its end is rotatably connected to the three-way pipe 12, and opposite external threads 2024 are provided on both sides of the guide tube 202, and the guide tube 202 is respectively threaded to two axial sliding plates 204, and a concave-convex structure is provided between the axial sliding plates 204 and the introduction shell 201 to prevent synchronous turnover. In this structure, the middle section of the guide tube 202 is integrated with the external thread 2024 and the guide port 2025, and the introduction shell 201 is provided with a limiting protrusion 2012 along its length direction, and the axial sliding plate 204 is provided with a limiting through groove 2041 along its length direction, and each axial sliding plate 204 is slidably connected to the limiting protrusion 2012 through the limiting through groove 2041.

[0100] Deformable capsules 207 are provided between the two ends of the radial expansion tube 206 and each caliber fixed tube. After the radial expansion tube 206 is reduced in diameter, the deformation capsule 207 is pressed inward to deform, thereby blocking the excess area of ​​each filter screen 205, and at the same time ensuring the sealing effect at the joint surface of the radial expansion tube 206 and the caliber fixed tube. In this structure, one end of the deformation capsule 207 is connected to the positioning point 2026 at the end of the caliber fixed tube, and an L-shaped arc-shaped pressing plate 2061 is provided on the second arc-shaped plate 2063. The inner portion of the L-shaped arc-shaped pressing plate 2061 is provided with a plurality of arc-shaped pressing plates 2061. The L-shaped arc-shaped pressing plate 2061 acts on the other end of the deformable capsule 207; when the radial telescopic tube 206 contracts, the L-shaped arc-shaped pressing plate 2061 acts on the deformable capsule 207, and the deformable capsule 207 is centered on the positioning point 2026, and the amount of gas inside it remains unchanged, so that the L-shaped arc-shaped pressing plate 2061 presses the deformable capsule 207 to converge toward the outer edge of the filter screen 205, and the conduction amount of the filter screen 205 becomes smaller, so that the diameter of the guide tube of the filter screen 205 is adapted to the reduced diameter of the L-shaped arc-shaped pressing plate 2061, and the bacterial agent is pressurized and transported in the liquid cavity 2021.

[0101] The specific implementation process is as follows: the bacterial agent is transported in the liquid chamber 2021 through the three-way pipe 12 into the fermentation kettle 1, and a section thereof passes through the annular deformable membrane 2065; the gas required for the bacterial agent passes through the air inlet 2011, and finally passes through the gas chamber 2022 and the three-way pipe 12 into the fermentation kettle 1; during this period, the air intake is controlled under the action of the axial sliding plate 204, and the gas acts on the annular deformable membrane 2065 from the outside to the inside during the process, so that the annular deformable membrane 2065 changes its diameter (becomes smaller), and then the problem of insufficient filling of the entire transport pipeline when processing a small batch of liquid bacterial agents is compensated by changing the passage diameter of the liquid chamber 2021.

[0102] In the description of the present invention, it should be understood that the terms "longitudinal", "lateral", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate orientations or positional relationships based on the orientations or positional relationships shown in the accompanying drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present invention. In the description of the present invention, unless otherwise specified and limited, it should be noted that the terms "installed", "connected" and "connected" should be understood in a broad sense, for example, it can be a mechanical connection or an electrical connection, or it can be the internal communication of two elements, it can be a direct connection, or it can be an indirect connection through an intermediate medium. For ordinary technicians in this field, the specific meanings of the above terms can be understood according to specific circumstances.

[0103] The above are only preferred embodiments of the present invention and are 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 in the protection scope of the present invention.

Claims

1. A device for preparing a composite microbial agent for aquaculture, characterized in that: include: A fermentation kettle (1) having a stirring assembly (3) vertically built therein, wherein a bacterial agent introduction assembly (2) and a sampling assembly (4) are respectively arranged above and below the fermentation kettle (1), and the output end of the bacterial agent introduction assembly (2) intersects with the stirring end of the rotating shaft of the stirring assembly (3) in the fermentation kettle (1) through a three-way pipe (12); The bottom of the fermentation kettle (1) is connected to a base (102) via a connecting column (101), and the sampling component (4) is arranged between the bottom of the fermentation kettle (1) and the base (102), which is specifically a rotation drive component (6) driven by a first motor (13), and an intrusive manual drive component (5) is installed at the rotation drive component (6), thereby realizing free switching between automatic sampling and manual sampling; The bottom of the fermentation kettle (1) is provided with a plurality of outlets equidistantly along its circumference, and the sampling assembly (4) is composed of a number of cylinders (401) equal to the number of the outlets, and the cylinders (401) are respectively arranged at each outlet; The upper and lower ends of the cylinder (401) are respectively provided with a first opening and closing member (402) and a second opening and closing member (403) of the same structure, wherein the first opening and closing member (402) is composed of an upper opening and closing plate (4021) hinged in a cross shape, and the second opening and closing member (403) is composed of a lower opening and closing plate (4023) hinged in a cross shape, and a return spring is provided at the hinge. A trigger plate (4022) is extended outward from any opening and closing plate, and the two opening and closing plates seal the end surface of the cylinder (401) in a closed state; A shell (10) is further provided between the bottom of the fermentation kettle (1) and the base (102), the bottom of the cylinder (401) is a sampling port, and the shell (10) is used to isolate each sampling port from the rotating drive assembly (6); The rotation drive assembly (6) comprises an internal gear ring (601) which is connected as a whole and has two layers, the outer circumferences of the two internal gear rings (601) are respectively connected with a first lever (604) and a second lever (605), the two levers are arranged in a staggered manner, and the second lever (605) and the first lever (604) are respectively in the same horizontal plane with the trigger plates (4022) of the first opening and closing member (402) and the second opening and closing member (403); The rotation drive assembly (6) further comprises a second rotating gear (603) connected to the output end of the first motor (13), a plurality of first rotating gears (602) are provided between the second rotating gear (603) and the inner gear ring (601) in a circumferentially meshed manner, and the thickness of the first rotating gear (602) is greater than the thickness of the inner gear ring (601); The bacterial agent introduction component (2) comprises an introduction shell (201) and a flow guide tube (202) rotatably arranged in the introduction shell (201); an air inlet (2011) is provided on the side of the introduction shell (201); a liquid cavity (2021) and a gas cavity (2022) are arranged in parallel along the length direction of the flow guide tube (202); the liquid cavity (2021) is axially connected and located at the center; and the gas cavity (2022) faces one side of the three-way tube (12) and is in a conductive state; The guide tube (202) is provided with a conducting port (2025) at a position corresponding to the air inlet (2011); the guide tube (202) is provided with an axial sliding plate (204) at the conducting port (2025) for controlling the aperture and air intake volume by movement; The two ends of the flow guide tube (202) are configured as fixed-diameter tubes, and the middle thereof is configured as a radial telescopic tube (206) with a variable diameter between the liquid chamber (2021) and the gas chamber (2022); the radial telescopic tube (206) is internally provided with an arc spring (2064) for automatic reset; a filter (205) is installed between the end of the radial telescopic tube (206) and the fixed-diameter tube; pressure sensors are installed in the front and rear fixed-diameter tubes; the radial telescopic tube (206) adjusts the diameter of the liquid chamber (2021) by changing the air intake volume of the conduction port (2025), thereby compensating for the insufficient pressure supply of the liquid chamber (2021).

2. The device for preparing a composite microbial agent for aquaculture according to claim 1, characterized in that: One end of the manual drive member (5) is rotatably connected to the base (102), and includes an integrated worm (501) and a hand wheel (503), both of which are rotatably connected to the translation block (502), and the worm (501) corresponds to any one of the first rotating gears (602). Two elastic limiting protrusions (103) are provided on the base (102), which correspond to the circular grooves at the bottom of the translation block (502) and are used to fix the two states of engagement and disengagement between the worm (501) and the first rotating gear (602).

3. The device for preparing a composite microbial agent for aquaculture according to claim 2, characterized in that: A second motor (203) is provided outside the introduction shell (201), the output end of which is meshed with a ring gear (2023) passing through one end of the guide tube (202) and extending out of the introduction shell (201) through a gear, and the end of the second motor (203) is rotatably connected to the three-way pipe (12); The two sides of the guide tube (202) are provided with opposite external threads (2024), the guide tube (202) is respectively threadedly connected to the two axial sliding plates (204), and a concave-convex structure is provided between the axial sliding plates (204) and the introduction shell (201) to prevent synchronous rotation; Deformable capsules (207) are provided between the two ends of the radial telescopic tube (206) and each fixed-caliber tube. After the radial telescopic tube (206) is reduced in diameter, the deformable capsule (207) is pressed inward to deform, thereby sealing the excess area of ​​each filter screen (205), while ensuring the sealing effect at the joint surface between the radial telescopic tube (206) and the fixed-caliber tube.

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