Low mortality pond complex microbial bacteria liquid into the pool of pond improvement process

Abstract

The present application relates to the field of aquaculture environment improvement technology, and particularly relates to a low mortality rate breeding pond composite microbial bacteria liquid pool entry pond starting improvement process, comprising: S100, preparation and activation of special composite microbial bacteria liquid: Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris and denitrifying bacteria are mixed according to a mass ratio of 2:1:3:2:2, sterile water containing molasses and amino acid polypeptide is added, and the mixture is micro-aerobically expanded and cultured at 28-30 DEG C for 24-36 hours to prepare a composite bacteria liquid with an effective total number of viable bacteria CFU / mL. The present application introduces a specific space-time coordination process at two high-risk nodes of fry pool entry and pond starting fishing, uses a gradual mixed liquid immersion bath regulated by temperature difference and osmotic pressure before pool entry, makes the fry adapt to the original pool water physicochemical indexes smoothly and attach the probiotic film on the body surface quickly, and reduces the acute stress mortality rate in the initial stage of pool entry.

Description

Technical Field

[0001] This invention relates to the field of aquaculture environment improvement technology, specifically to a process for improving aquaculture ponds by introducing compound microbial inoculants into the pond and then removing the pond. Background Technology

[0002] In modern aquaculture, the addition of compound microbial preparations to regulate water and bottom sediment has become a routine operation. However, current microbial application methods mainly involve periodic whole-pond application during the aquaculture cycle, failing to provide targeted and systematic intervention processes for the two specific stages with the highest mortality rates in aquatic animal farming—"seedling introduction" and "harvesting"—leading to the following two core problems in actual production: First, there is the issue of acute stress and infection during the seedling introduction stage. During the transfer from the transport water to the rearing pond, seedlings face sudden changes in water temperature and inorganic salt osmotic pressure, making them highly susceptible to severe physiological stress. Current techniques typically only involve pre-sprinkling bacterial solutions into the pond water, failing to provide pre-emptive physiochemical buffering for individual seedlings. After introduction, under the stress-induced state of weakened immunity, the seedlings lack the protection of dominant beneficial bacteria on their skin and intestinal tract, making them highly vulnerable to infection by native opportunistic pathogens in the rearing pond, thus triggering large-scale mortality in the early stages of introduction.

[0003] Secondly, there is the problem of acute poisoning caused by the disturbance of bottom sediment during the pond harvesting stage. In the middle and late stages of aquaculture, a large amount of uneaten feed and feces accumulate at the bottom of the pond, forming a reducing sediment rich in harmful substances such as hydrogen sulfide and ammonia nitrogen. During harvesting operations, mechanical actions such as netting violently agitate the bottom sediment, causing a large amount of toxic substances trapped at the bottom to be released into the water instantly. Conventional liquid microbial spraying processes, because the bacterial solution is mostly suspended in the middle and upper water layers, cannot penetrate the water layer to form a high-concentration micro-ecological degradation layer on the surface of the bottom sediment, making it difficult to eliminate bottom sediment toxins in advance. This directly leads to extremely high mortality and disease rates in aquatic animals under the dual effects of pond harvesting stress and acute toxic substance impact. Summary of the Invention

[0004] To achieve the above objectives, the present invention provides the following technical solution: This invention provides a process for improving the inoculation and removal of aquaculture ponds with a low mortality rate using a compound microbial inoculum solution, comprising the following steps: S100, Preparation and activation of the special compound microbial inoculum solution: Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris, and denitrifying bacteria are mixed in a mass ratio of 2:1:3:2:2, added to sterile water containing molasses and amino acid peptides, and cultured under micro-aeration at 28-30℃ for 24-36 hours to obtain the total effective viable bacteria count. CFU / mL compound bacterial solution; S200. Pre-planting in the pond: 3-5 days before the seedlings are introduced into the pond, spray the aforementioned compound bacterial solution into the pond at a rate of 1.5-2.0 L / mu·m, and simultaneously turn on the bottom micro-pore aeration to maintain the dissolved oxygen level in the water. mg / L; S300. Soaking of seedlings before entering the pond: On the day the seedlings are placed in the pond, soak them in a mixture of the compound bacterial solution diluted 50-100 times and 0.1% sodium vitamin C for 15-20 minutes before transferring them to the breeding pond. S400, Pond Improvement Before Harvesting: 10 days before the expected harvest, use potassium persulfate compound salt to oxidize and improve the pond bottom; 48 hours after the bottom improvement, mix the compound bacterial solution with zeolite powder at a mass ratio of 1:5 and then sprinkle it onto the bottom of the pond, using 3.0-4.0L of bacterial solution / mu·m; 22 days before harvesting, stop feeding conventional feed and switch to feed mixed with compound bacterial solution and immune polysaccharides.

[0005] Further, in step S100, the initial effective viable count of the single-strain powders of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris, and denitrifying bacteria before mixing is not less than 1.0 × 10^8 CFU / g; The specific preparation parameters for the sterile water containing molasses and amino acid peptides are as follows: based on the total mass of sterile water as 100%, the amount of molasses added is 3.0%-5.0%, the amount of amino acid peptides added is 0.4%-0.6%, and the initial pH value of the sterile water is adjusted to 6.8-7.2.

[0006] Furthermore, the specific dynamic control operation of the "micro-oxygenation expansion culture" in step S100 is as follows: Under constant temperature conditions of 28-30℃, microporous aeration is carried out in the culture to maintain the dissolved oxygen concentration of the fermentation broth at 1.5-2.5 mg / L; and during the expansion culture, mechanical stirring is started every 4-6 hours, with a stirring speed of 100-150 r / min and a single stirring duration of 10-15 minutes.

[0007] Furthermore, in step S200, the operation time for spraying the compound bacterial solution is limited to 8:00-10:00 am on a sunny day, and before spraying, the compound bacterial solution is mixed with the original pond water in the aquaculture pond at a volume ratio of 1:20-1:50 and then evenly sprayed throughout the pond. In step S200, the bottom microporous aeration adopts a disc-type microporous aerator. The arrangement density of the disc-type microporous aerator is 3-5 per acre, and the installation height is 15-20cm away from the surface of the mud at the bottom of the aquaculture pond.

[0008] Furthermore, in step S200, after spraying the compound bacterial solution and opening the bottom micropores for aeration, the apparent indicators of the water body need to be monitored. When the water transparency of the aquaculture pond is stable at 25-35cm and the water color is yellow-green or tea-brown, the water body pre-planting is completed.

[0009] Furthermore, in step S300, the temperature difference between the mixture and the water used for seedling transfer is controlled within a certain range. Within; and during the immersion period, intermittent micro-oxygenation is continuously performed on the mixture to maintain the dissolved oxygen concentration of the mixture at 6.0-8.0 mg / L.

[0010] Furthermore, in step S300, the water source used to dilute the compound bacterial solution is the surface water of the aquaculture pond; and in the mixture, glucose with a mass fraction of 0.2%-0.5% is additionally added as an inorganic salt osmotic pressure regulator.

[0011] Furthermore, in step S300, the specific operation of "transferring to the aquaculture pond" after immersion is as follows: After the set immersion time is reached, the original pond water of the breeding pond is gradually introduced into the immersion container containing the mixture at a flow rate of 5-10 L / min until the volume ratio of the original pond water in the mixture reaches 70%-80%. Then, the water introduction is stopped, and the mixture is left to stand for 5-10 minutes before the seedlings are released into the breeding pond with the wind.

[0012] Furthermore, in step S400, the potassium persulfate compound salt used is a specially prepared bottom sedimentation slow-release tablet with an effective active oxygen mass fraction of not less than 10%, and the amount of scattering is controlled at 1.0-1.5 kg / mu, and the scattering area is mainly concentrated in the feeding area and deep water sewage discharge area of ​​the aquaculture pond. In step S400, the particle size of the zeolite powder is limited to 60-80 mesh; the specific operation process of the "mixed adsorption" is as follows: the composite bacterial liquid is evenly sprayed onto the surface of the zeolite powder in the form of high pressure micro-mist, while spraying and stirring, and then left to stand and solidify in a cool, dark place for 2-3 hours. After the surface is free of water and has a loose granular shape, the bottom is then sprinkled.

[0013] Further, in step S400, the specific preparation process of the feed mixed with the compound bacterial solution and immunopolysaccharide is as follows: based on the mass of the conventional feed as 100%, 2.0% of the compound bacterial solution and 0.3%-0.5% of yeast β-glucan are added sequentially as immunopolysaccharides. After thorough mixing and absorption, 1.0%-1.5% of refined fish oil is used to spray a coating on the surface of the feed.

[0014] Beneficial effects This invention introduces specific spatiotemporal synergistic processes at two high-risk stages: seedling introduction into the pond and harvesting. Before introduction, a gradual mixed-liquid immersion process with temperature and osmotic pressure control is used to allow seedlings to smoothly adapt to the original pond water's physicochemical properties and rapidly attach a probiotic film to their body surface, reducing acute stress mortality in the early stages of introduction. Simultaneously, before harvesting, a bottom-targeted oxidation combined with a sedimentation and solidification process using zeolite powder to adsorb bacterial solution is employed to pre-construct a microecological degradation and physical sealing layer on the bottom sediment surface. Combined with targeted feeding of high-energy coated immune feed, this effectively blocks the release of bottom toxins and physiological damage caused by mechanical agitation during harvesting. Without relying on large amounts of chemical antibiotics, this significantly improves the overall survival rate of aquatic animals at critical transplanting stages. Attached Figure Description

[0015] Figure 1 This is a flowchart of the overall process of the present invention; Figure 2 This is a detailed process flow diagram of step S100 of the present invention; Figure 3 This is a detailed process flow diagram of step S200 of the present invention; Figure 4 This is a detailed process flow diagram of step S300 of the present invention; Detailed Implementation

[0016] To enable those skilled in the art to better understand the present invention, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. 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 should fall within the scope of protection of the present invention.

[0017] It should be noted that the terms "first," "second," etc., in the specification, claims, and accompanying drawings of this invention are used to distinguish similar objects and are not necessarily used to describe a specific order or sequence. It should be understood that such data can be interchanged where appropriate so that the embodiments of the invention described herein can be implemented in orders other than those illustrated or described herein. Furthermore, the terms "comprising" and "having," and any variations thereof, are intended to cover a non-exclusive inclusion; for example, a process, method, system, product, or apparatus that comprises a series of steps or units is not necessarily limited to those steps or units explicitly listed, but includes other steps or units not explicitly listed or inherent to such processes, methods, products, or apparatus.

[0018] The present invention will now be described in further detail with reference to the accompanying drawings: like Figures 1-4As shown, the process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum solution into the pond includes the following steps: Preparation and activation of S100 special compound microbial culture: Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris, and denitrifying bacteria are mixed in a mass ratio of 2:1:3:2:2, added to sterile water containing molasses and amino acid peptides, and cultured under micro-aerobic conditions at 28-30℃ for 24-36 hours to obtain the total effective viable bacteria count. CFU / mL compound bacterial solution; S200, Pre-planting in the pond before stocking: 3-5 days before stocking, spray compound bacterial solution into the pond at a rate of 1.5-2.0 L / mu·m, and simultaneously turn on the bottom micro-pore aeration to maintain dissolved oxygen levels in the water. mg / L; S300. Soaking of seedlings before entering the pond: On the day the seedlings are placed in the pond, soak them in a mixture of compound bacterial solution diluted 50-100 times and 0.1% sodium vitamin C for 15-20 minutes before transferring them to the breeding pond. S400, Pond Improvement Before Harvesting: 10 days before the expected harvest, use potassium persulfate compound salt to oxidize and improve the pond bottom; 48 hours after the bottom improvement, mix compound bacterial solution and zeolite powder at a mass ratio of 1:5 and then sprinkle it onto the bottom of the pond, using 3.0-4.0L of bacterial solution / acre·meter; 22 days before harvesting, stop feeding conventional feed and switch to feed mixed with compound bacterial solution and immune polysaccharides.

[0019] Furthermore, the specific implementation process of step S100 is as follows: Step S100 involves the preparation and activation of the specialized composite microbial culture solution. Step S100 specifically includes four consecutive process stages: weighing raw materials, preparing the basic culture medium, inoculation and mixing, and dynamic parameter expansion culture.

[0020] First, the raw materials were weighed. Bacillus subtilis powder, Bacillus licheniformis powder, Lactobacillus plantarum powder, Rhodopseudomonas palustris powder, and denitrifying bacteria powder were obtained separately. The initial viable count of each of the five single-strain powders was not less than 1.0 × 10^8 CFU / g. Using a high-precision electronic balance, the Bacillus subtilis powder, Bacillus licheniformis powder, Lactobacillus plantarum powder, Rhodopseudomonas palustris powder, and denitrifying bacteria powder were accurately weighed at a mass ratio of 2:1:3:2:2 and placed in sterile containers for later use.

[0021] The basic culture medium was then prepared. Using sterile water as the total mass (100%), sterile water was added to the fermenter. Then, molasses (3.0% to 5.0% by mass) and amino acid peptides (0.4% to 0.6% by mass) were added sequentially to the sterile water. The fermenter's built-in agitator was activated, and stirring was maintained until the molasses and amino acid peptides were completely dissolved and homogeneous in the sterile water, forming a liquid basic culture medium. The initial pH of the basic culture medium was adjusted and stabilized within the range of 6.8 to 7.2 using a 0.1 mol / L sodium hydroxide solution or hydrochloric acid solution.

[0022] Next, the inoculation, mixing, and dynamic parameter expansion operations are performed. The five types of bacterial powder prepared in a sterile container are added to the pH-adjusted basal culture medium. The temperature control system of the fermenter is turned on, setting the constant temperature within the tank to between 28°C and 30°C. Simultaneously, the microporous aeration device connected to the bottom of the fermenter is started, and the dissolved oxygen concentration inside the fermentation broth is strictly maintained between 1.5 mg / L and 2.5 mg / L by adjusting the opening of the air inlet valve. During the continuous culture period of 24 to 36 hours, intermittent mechanical stirring is performed through an automated control program: the fermenter's stirring motor is started every 4 to 6 hours, controlling the speed of the stirring paddle to be constant at 100 r / min to 150 r / min, with each stirring action lasting 10 to 15 minutes before automatically stopping. Intermittent stirring and constant microporous aeration are alternated until the set total culture time is reached.

[0023] After the culture cycle is completed, the activated fermentation broth sample is extracted from the sampling port of the fermenter for microplate counting. When the total number of viable bacteria in the fermentation broth reaches or exceeds 2×10^9 CFU / mL, temperature control and aeration are stopped, and the preparation and activation of the special compound microbial culture solution are completed.

[0024] To further illustrate the specific implementation details of step S100 under different parameter conditions, three specific point value examples for S100 are provided below: Example 1 (Example of lower limit parameter value): Prepare 200g of Bacillus subtilis powder, 100g of Bacillus licheniformis powder, 300g of Lactobacillus plantarum powder, 200g of Rhodopseudomonas palustris powder, and 200g of denitrifying bacteria powder, each with an initial viable count of 1.0 × 10^8 CFU / g. Add 100kg of sterile water to the fermenter, then add 3.0kg of molasses and 0.4kg of amino acid peptides sequentially. After stirring and dissolving, adjust the initial pH of the basal culture medium to 6.8 using hydrochloric acid solution. Add 1kg of the prepared bacterial powder to the basal culture medium. Set the fermenter temperature to 28℃ and start the microporous aeration device to maintain a constant dissolved oxygen concentration of 1.5mg / L. During the 24-hour culture period, start the stirring motor every 4 hours, stirring continuously at 100r / min for 10 minutes. After 24 hours, take samples for testing; the total viable count was found to be 2.1 × 10^9 CFU / mL, thus obtaining the composite microbial culture.

[0025] Example 2 (Example of optimal parameter centering): Prepare 400g of Bacillus subtilis powder, 200g of Bacillus licheniformis powder, 600g of Lactobacillus plantarum powder, 400g of Rhodopseudomonas palustris powder, and 400g of denitrifying bacteria powder, all with an initial effective viable count of 1.0 × 10^8 CFU / g. Add 200kg of sterile water to the fermenter, then add 8.0kg of molasses and 1.0kg of amino acid peptides sequentially. After stirring and dissolving, adjust the initial pH of the basal culture medium to 7.0 using sodium hydroxide solution. Add a total of 2kg of the prepared bacterial powder to the basal culture medium. Set the fermenter temperature to 29℃ and start the microporous aeration device to maintain a constant dissolved oxygen concentration of 2.0mg / L. During the 30-hour culture period, start the stirring motor every 5 hours, stirring continuously at 120r / min for 12 minutes. After 30 hours, take samples for testing; the total effective viable count was found to be 3.5 × 10^9 CFU / mL, thus obtaining the composite microbial culture.

[0026] Example 3 (Example of upper limit parameter value): Prepare 200g of Bacillus subtilis powder, 100g of Bacillus licheniformis powder, 300g of Lactobacillus plantarum powder, 200g of Rhodopseudomonas palustris powder, and 200g of denitrifying bacteria powder, all with an initial viable count of 1.0 × 10^8 CFU / g. Add 100kg of sterile water to the fermenter, then add 5.0kg of molasses and 0.6kg of amino acid peptides sequentially. After stirring and dissolving, adjust the initial pH of the basal culture medium to 7.2 using sodium hydroxide solution. Add 1kg of the prepared bacterial powder to the basal culture medium. Set the fermenter temperature to 30℃ and start the microporous aeration device to maintain a constant dissolved oxygen concentration of 2.5mg / L. During the 36-hour culture period, start the stirring motor every 6 hours, stirring continuously at 150r / min for 15 minutes. After 36 hours, take samples for testing; the total viable count was found to be 2.8 × 10^9 CFU / mL, thus obtaining the composite microbial culture.

[0027] Furthermore, the specific implementation process of step S200 is as follows: Step S200 is the pre-planting operation of the water body before entering the pool. Step S200 specifically includes four continuous process stages: aeration equipment layout, in-situ dilution of bacterial solution, timed and quantitative spraying, and water quality index monitoring.

[0028] First, the aeration equipment should be installed. Three to five days before the seedlings are released into the aquaculture pond, disc-type microporous aerators should be installed at the bottom of the pond. The density of the disc-type microporous aerators should be controlled at 3 to 5 per acre. During installation, the installation height of the disc-type microporous aerators should be fixed at 15cm to 20cm above the surface of the bottom mud using fixed brackets to prevent the airflow from directly agitating the bottom mud surface. After installation, the bottom microporous aeration system should be turned on for continuous aeration to maintain a stable dissolved oxygen concentration in the pond water above 5.0mg / L.

[0029] Following this, in-situ dilution of the bacterial solution and timed, quantitative application were performed. Application was carried out between 8:00 AM and 10:00 AM on a sunny day. The area and water depth of the aquaculture pond were calculated, and the compound microbial solution prepared in step S100 was taken at a dosage of 1.5 L / mu·m to 2.0 L / mu·m. The extracted compound microbial solution was poured into a dilution container containing the original pond water, controlling the volume ratio of the compound microbial solution to the original pond water to be 1:20 to 1:50. The two were thoroughly mixed manually or mechanically to prepare the diluted bacterial solution. The diluted bacterial solution was then evenly sprayed onto the surface of the aquaculture pond water using a spraying device.

[0030] Finally, water quality monitoring was performed. During the application of the compound microbial inoculant and the continuous operation of the bottom microporous aeration system, the water transparency was measured at fixed times daily using a Sesquimet disc, and water color changes were observed simultaneously. Monitoring continued until the water transparency in the aquaculture pond remained stably within the range of 25cm to 35cm, and the water color appeared yellowish-green or brownish-yellow. At this point, the pre-planting stage was considered complete, and the subsequent seedling introduction process could begin.

[0031] To further illustrate the specific implementation details of step S200 under different parameter conditions, three specific point value examples for S200 are provided below: Example 4 (Example of lower limit parameter values): Three days before stocking the seedlings, pre-planting was carried out in a 10-mu (approximately 6.7 hectares) aquaculture pond with a water depth of 1.5 meters. Disc-type microporous aerators were installed at the bottom of the pond, with a density of 3 per mu (approximately 0.067 hectares) and an installation height of 15 cm from the bottom sediment surface. The aeration system was activated, and the gas flow rate was adjusted to maintain the dissolved oxygen concentration in the water at 5.0 mg / L. At 8:00 AM on a sunny day, 22.5 L (equivalent to 1.5 L / mu·m) of the activated compound microbial solution from step S100 was taken. The solution was added to a dilution tank, along with 450 L of original pond water (volume ratio 1:20), stirred thoroughly, and then sprayed throughout the pond. Aeration was continued, and monitoring was conducted daily. On the third day after spraying, when the water transparency reached 25 cm and the water color turned yellowish-green, the pre-planting was completed.

[0032] Example 5 (Example of optimal parameter centering): Four days before the seedlings were introduced into the pond, pre-planting was carried out in a 20-mu (approximately 3.3 hectares) aquaculture pond with a water depth of 2.0 meters. Disc-type microporous aerators were installed at the bottom of the pond, with a density of 4 per mu (approximately 0.067 hectares) and an installation height of 18 cm from the bottom sediment surface. The aeration system was started, and the gas flow rate was adjusted to maintain the dissolved oxygen concentration in the water at 5.5 mg / L. At 9:00 AM on a sunny day, 72 L (equivalent to 1.8 L / mu·m) of the activated compound microbial solution from step S100 was taken. The solution was added to a dilution tank, along with 2520 L of original pond water (volume ratio 1:35), stirred thoroughly, and then sprayed throughout the pond. Aeration was continued, and monitoring was conducted daily. On the fourth day after spraying, when the water transparency reached 30 cm and the water color turned a tea-brown, the pre-planting was completed.

[0033] Example 6 (Example of upper limit parameter value): Five days before the seedlings were introduced into the pond, pre-planting was carried out in a 15-mu (approximately 1 hectare) aquaculture pond with a water depth of 1.8 meters. Disc-type microporous aerators were installed at the bottom of the pond, with a density of 5 per mu (approximately 0.067 hectares) and an installation height of 20 cm from the bottom sediment surface. The aeration system was started, and the gas flow rate was adjusted to maintain the dissolved oxygen concentration in the water at 6.2 mg / L. At 10:00 AM on a sunny day, 54 L (equivalent to 2.0 L / mu·m) of the activated compound microbial solution from step S100 was taken. The solution was added to a dilution tank, along with 2700 L of original pond water (volume ratio 1:50), stirred thoroughly, and then sprayed throughout the pond. Aeration was continued, and monitoring was conducted daily. On the fourth day after spraying, when the water transparency reached 35 cm and the water color turned yellowish-green, the pre-planting was completed.

[0034] Furthermore, the specific implementation process of step S300 is as follows: Step S300 is the soaking operation before the seedlings are placed in the pond. Step S300 specifically includes four continuous process stages: in-situ preparation of the soaking solution, temperature and dissolved oxygen control, gradual adaptation to raw water, and downstream release.

[0035] First, the in-situ preparation of the bath solution is carried out. On the day the seedlings are released into the pond, surface water from the aquaculture pond is drawn as a dilution source and injected into the bath container. The compound microbial solution prepared in step S100 is added to the bath container at a volume dilution ratio of 50 to 100 times. Simultaneously, sodium vitamin C and glucose (as an inorganic salt osmotic pressure regulator) are added to the bath container. Based on the total mass of the water in the bath container (100%), the amount of sodium vitamin C added is controlled at 0.1%, and the amount of glucose added is controlled at 0.2% to 0.5%. The above components are completely dispersed and dissolved evenly using a stirring tool to prepare the bath mixture.

[0036] Temperature and dissolved oxygen control were then implemented. Before transferring the seedlings into the immersion container, the temperatures of the water used for seedling transport and the immersion solution were measured using a thermometer. The temperature difference between the immersion solution and the water used for seedling transport was strictly controlled within 1.0℃ by adding ice to the outer wall of the immersion container for cooling or adding warm water for temperature adjustment. After temperature adjustment, the seedlings were immersed in the immersion solution, and the total immersion time was set to 15 to 20 minutes. During continuous immersion, an oxygenation pump was activated in conjunction with sand strips to intermittently provide micro-oxygenation to the immersion solution. The gas flow rate was adjusted in real-time using an online dissolved oxygen meter to maintain a constant dissolved oxygen concentration of 6.0 mg / L to 8.0 mg / L within the immersion solution.

[0037] Next, a gradual acclimatization process will be implemented. After the immersion period has reached the specified duration, the seedlings will remain in the immersion container. A metered-flow pump connected to the rearing pond will be turned on, gradually introducing the original pond water into the immersion container at a fixed flow rate of 5 L / min to 10 L / min. During the introduction of the original pond water, the liquid level and volume changes in the container will be observed. When the introduced original pond water volume accounts for 70% to 80% of the total volume of the mixed liquid in the immersion container, the metered-flow pump will be turned off to stop the water introduction. After stopping the water introduction, the seedlings will be allowed to stand in the container with the mixed water for 5 to 10 minutes to allow them to gradually adapt to the physical and chemical properties of the original pond water.

[0038] Finally, the downwind release operation is carried out. After the settling and buffering period, the container containing the seedlings and the mixed solution is transported to the downwind side of the aquaculture pond, and the seedlings are slowly poured into the aquaculture pond in the direction of the prevailing wind, completing the entire pond release process.

[0039] To further illustrate the specific implementation details of step S300 under different parameter conditions, three specific point value examples for S300 are provided below: Example 7 (Example of lower limit parameter values): On the day the Pacific white shrimp larvae were introduced into the pond, 500L of surface water from the rearing pond was drawn and injected into the bathing tank. 10L of the activated compound microbial solution from step S100 (diluted 50 times by volume), 0.51kg of sodium vitamin C (0.1% by mass), and 1.02kg of glucose (0.2% by mass) were added and stirred thoroughly to prepare the bathing mixture. The temperature of the water used for shrimp larvae transfer was measured at 26.5℃. The temperature of the bathing mixture was stabilized at 25.5℃ (temperature difference of -1.0℃) through physical temperature regulation. The shrimp larvae were immersed in the bathing mixture for 15 minutes, during which the oxygen pump was used intermittently to maintain a dissolved oxygen concentration of 6.0mg / L. After 15 minutes of immersion, the original pond water was injected into the bathing tank at a flow rate of 5L / min until the injected original pond water volume reached 1200L (70% of the total volume of the mixture), at which point the water injection was stopped. After letting the shrimp larvae sit in the bucket for 5 minutes, transport them to the downwind side of the pond and release them into the pond with the wind at your back.

[0040] Example 8 (Example of optimal parameter center value): On the day the California bass fry were introduced into the pond, 750L of surface water from the rearing pond was pumped into the bathing tank. 10L of the activated compound microbial solution (diluted 75 times by volume) from step S100, 0.76kg of sodium vitamin C (0.1% by mass), and 2.66kg of glucose (0.35% by mass) were added and stirred thoroughly to prepare the bathing mixture. The temperature of the water used for fry transfer was measured at 22.0℃, and the temperature of the bathing mixture was stabilized at 22.2℃ (temperature difference of 0.2℃) by temperature adjustment. The fry were immersed in the bathing mixture for 18 minutes, during which the oxygen pump was used intermittently to maintain a dissolved oxygen concentration of 7.0mg / L. After 18 minutes of immersion, the water pump was started to inject water from the original rearing pond into the tank at a flow rate of 8L / min until the injected water volume reached 2280L (75% of the total volume of the mixture), at which point the water injection was stopped. After letting the fish fry sit in the tank for 8 minutes, transport them to the downwind side of the pond and release them into the pond downwind.

[0041] Example 9 (Example of upper limit parameter value): On the day the Chinese mitten crab larvae were introduced into the pond, 1000L of surface water from the rearing pond was pumped into the bathing tank. 10L of the activated compound microbial solution (diluted 100 times by volume) from step S100, 1.01kg of sodium vitamin C (0.1% by mass), and 5.05kg of glucose (0.5% by mass) were added and stirred thoroughly to prepare the bathing mixture. The temperature of the water used for transporting the larvae was measured at 18.0℃. The temperature of the bathing mixture was stabilized at 19.0℃ (temperature difference of +1.0℃) by temperature adjustment. The larvae were immersed in the bathing mixture for 20 minutes, during which the oxygen pump was used intermittently to maintain a dissolved oxygen concentration of 8.0mg / L. After 20 minutes of immersion, the original pond water was pumped into the bathing tank at a flow rate of 10L / min until the volume of the injected original pond water reached 4040L (80% of the total volume of the mixture), at which point the water injection was stopped. After letting the crab seedlings sit in the pond for 10 minutes, transport them to the downwind side of the pond and release them into the pond downwind.

[0042] Furthermore, the specific implementation process of step S400 is as follows: Step S400 is the pre-pond improvement operation. Step S400 specifically includes three continuous process stages: bottom sediment targeted oxidation, microparticle adsorption and solidification and spreading, and coated feed preparation and feeding.

[0043] First, targeted oxidation of the bottom sediment is carried out. Ten days before the expected harvesting operation, potassium persulfate compound salt bottom sedimentation slow-release tablets with an effective active oxygen mass fraction of not less than 10% are obtained. The tablets are accurately weighed according to a dosage standard of 1.0 kg / mu to 1.5 kg / mu. The weighed potassium persulfate compound salt bottom sedimentation slow-release tablets are then directly scattered into the aquaculture pond water. During the scattering operation, the scattering points are controlled to concentrate in the feeding area and deep-water drainage area within the aquaculture pond.

[0044] Following this, particle adsorption solidification and scattering operations were performed. 48 hours after the bottom sediment targeted oxidation operation, zeolite powder with a particle size of 60 to 80 mesh was obtained as a carrier. The composite microbial inoculum and zeolite powder prepared in step S100 were weighed out at a mass ratio of 1:5. A high-pressure micro-mist spraying device was activated to uniformly spray the weighed composite microbial inoculum onto the surface of the zeolite powder in a high-pressure micro-mist fluid form. Simultaneously with spraying the composite microbial inoculum, a mechanical stirring device was activated to continuously stir the zeolite powder. After spraying and stirring, the mixture was transferred to a cool, dark environment and allowed to solidify for 2 to 3 hours. Solidification was stopped when there was no visible water on the surface of the mixture and it exhibited a loose granular form. Following a dosage conversion standard of 3.0 L / mu·m to 4.0 L / mu·m, the solidified loose granular mixture was scattered throughout the aquaculture pond, allowing it to naturally settle to the bottom sediment surface.

[0045] Finally, the coated feed preparation and feeding operations are carried out. Two days before the expected harvest, feeding of conventional feed into the aquaculture ponds is stopped. Taking the total mass of the conventional feed for a single feeding as 100%, 2.0% by mass of the compound microbial inoculum prepared in step S100 and 0.3% to 0.5% by mass of yeast β-glucan are added sequentially to the conventional feed. The mixer is turned on and thoroughly mixed until the liquid and powder are completely absorbed by the conventional feed. After absorption, 1.0% to 1.5% by mass of refined fish oil is sprayed onto the surface of the mixed feed using an atomizing nozzle to complete the surface coating treatment. The coated feed is then fed into the aquaculture ponds at the usual feeding rate.

[0046] To further illustrate the specific implementation details of step S400 under different parameter conditions, three specific point value examples for S400 are provided below: Example 10 (Example of lower limit parameter values): Ten days before the expected harvest of Litopenaeus vannamei shrimp, obtain potassium persulfate compound salt bottom sedimentation slow-release tablets with an effective active oxygen mass fraction of 10%. Apply 1.0 kg / mu (total 10.0 kg) to a 10-mu (approximately 6.7 hectares) pond, focusing on the feeding area and deep-water drainage area. Forty-eight hours after application, take 150.0 kg of 60-mesh zeolite powder and 30.0 kg (approximately 30 L, corresponding to a dosage of 3.0 L / mu·m for a 10-mu pond with a water depth of 1 meter, a ratio of 1:5) of the activated compound bacterial solution from step S100. Spray the bacterial solution onto the zeolite powder surface using high-pressure micro-mist and continuously stir. Let it stand in a cool, dark place for 2 hours to solidify until it becomes loose granules before spreading it into the pond. Two days before harvesting, stop feeding regular feed. Take 100.0 kg of regular feed, add 2.0 kg of compound bacterial solution and 0.3 kg of yeast β-glucan, stir and absorb, then spray 1.0 kg of refined fish oil on the surface for coating treatment, and then put it into the breeding pond.

[0047] Example 11 (Example of optimal parameter centering value): Ten days before the expected harvest of California bass, potassium persulfate compound salt bottom sedimentation slow-release tablets with an effective active oxygen mass fraction of 12% were obtained and concentratedly scattered in the feeding area and deep-water drainage area of ​​a 20-acre aquaculture pond at a dosage of 1.25 kg / acre (totaling 25.0 kg). 48 hours after scattering, 525.0 kg of 70-mesh zeolite powder and 105.0 kg of compound bacterial solution activated in step S100 were taken (approximately 105 L in volume, corresponding to a dosage of 3.5 L / acre·meter for a water depth of 1.5 meters on 20 acres, a ratio of 1:5). The bacterial solution was sprayed onto the surface of the zeolite powder in the form of high-pressure micro-mist and continuously stirred. It was then left to solidify in a cool, dark place for 2.5 hours until it became loose granules before being scattered into the pond. Two days before harvesting, stop feeding regular feed. Take 200.0 kg of regular feed, add 4.0 kg of compound bacterial solution and 0.8 kg of yeast β-glucan, stir and absorb, then spray 2.5 kg of refined fish oil on the surface for coating treatment, and then put it into the breeding pond.

[0048] Example 12 (Example of upper limit parameter value): Ten days before the expected harvest of Chinese mitten crabs, obtain potassium persulfate compound salt bottom sedimentation slow-release tablets with an effective active oxygen mass fraction of 15%. Apply 1.5 kg / mu (totaling 22.5 kg) to the feeding area and deep-water drainage area of ​​a 15-mu (approximately 1 hectare) aquaculture pond. Forty-eight hours after application, take 540.0 kg of 80-mesh zeolite powder and 108.0 kg of the compound bacterial solution activated in step S100 (approximately 108 L, corresponding to a dosage of 4.0 L / mu·m for a 15-mu water depth of 1.8 meters, a ratio of 1:5). Spray the bacterial solution onto the surface of the zeolite powder using high-pressure micro-mist and continuously stir. Let it stand in a cool, dark place for 3 hours to solidify until it becomes loose granules before spreading it into the pond. Two days before harvesting, stop feeding regular feed. Take 150.0 kg of regular feed, add 3.0 kg of compound bacterial solution and 0.75 kg of yeast β-glucan, stir and absorb, then spray 2.25 kg of refined fish oil on the surface for coating treatment, and then put it into the breeding pond.

[0049] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions will not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A process for improving the inoculation and removal of aquaculture ponds using compound microbial inoculants to reduce mortality rates, characterized by: Includes the following steps: Preparation and activation of S100 special compound microbial culture: Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris, and denitrifying bacteria are mixed in a mass ratio of 2:1:3:2:2, added to sterile water containing molasses and amino acid peptides, and cultured under micro-aerobic conditions at 28-30℃ for 24-36 hours to obtain the total effective viable bacteria count. CFU / mL compound bacterial solution; S200. Pre-planting in the pond: 3-5 days before the seedlings are introduced into the pond, spray the aforementioned compound bacterial solution into the pond at a rate of 1.5-2.0 L / mu·m, and simultaneously turn on the bottom micro-pore aeration to maintain the dissolved oxygen level in the water. mg / L; S300. Soaking of seedlings before entering the pond: On the day the seedlings are placed in the pond, soak them in a mixture of the compound bacterial solution diluted 50-100 times and 0.1% sodium vitamin C for 15-20 minutes before transferring them to the breeding pond. S400, Pond Improvement Before Harvesting: 10 days before the expected harvest, use potassium persulfate compound salt to oxidize and improve the pond bottom; 48 hours after the bottom improvement, mix the compound bacterial solution with zeolite powder at a mass ratio of 1:5 and then sprinkle it onto the bottom of the pond, using 3.0-4.0L of bacterial solution / mu·m; 22 days before harvesting, stop feeding conventional feed and switch to feed mixed with compound bacterial solution and immune polysaccharides.

2. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond, as described in claim 1, is characterized in that... In step S100, the initial effective viable count of the single-strain powders of Bacillus subtilis, Bacillus licheniformis, Lactobacillus plantarum, Rhodopseudomonas palustris, and denitrifying bacteria before mixing is not less than 1.0 × 10^8 CFU / g. The specific preparation parameters for the sterile water containing molasses and amino acid peptides are as follows: based on the total mass of sterile water as 100%, the amount of molasses added is 3.0%-5.0%, the amount of amino acid peptides added is 0.4%-0.6%, and the initial pH value of the sterile water is adjusted to 6.8-7.

2.

3. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond, as described in claim 2, is characterized in that... The specific dynamic control operation of the "micro-oxygenated expansion culture" in step S100 is as follows: Under constant temperature conditions of 28-30℃, microporous aeration is carried out in the culture to maintain the dissolved oxygen concentration of the fermentation broth at 1.5-2.5 mg / L; and during the expansion culture, mechanical stirring is started every 4-6 hours, with a stirring speed of 100-150 r / min and a single stirring duration of 10-15 minutes.

4. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond, as described in claim 3, is characterized in that... In step S200, the operation time for spraying the compound bacterial solution is limited to 8:00-10:00 am on a sunny day. Before spraying, the compound bacterial solution is mixed with the original pond water in the aquaculture pond at a volume ratio of 1:20-1:50 and then evenly sprayed throughout the pond. In step S200, the bottom microporous aeration adopts a disc-type microporous aerator. The arrangement density of the disc-type microporous aerator is 3-5 per acre, and the installation height is 15-20cm away from the surface of the mud at the bottom of the aquaculture pond.

5. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum solution into the pond and removing the fish from the pond according to claim 4, is characterized in that, In step S200, after spraying the compound bacterial solution and opening the bottom micropores for aeration, the apparent indicators of the water body need to be monitored. When the water transparency of the aquaculture pond is stable at 25-35cm and the water color is yellow-green or tea-brown, the water body pre-planting is completed.

6. The process for improving low-mortality aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond according to claim 5, is characterized in that, In step S300, the temperature difference between the mixture and the water used for seedling transfer is controlled at... Within; and during the immersion period, intermittent micro-oxygenation is continuously performed on the mixture to maintain the dissolved oxygen concentration of the mixture at 6.0-8.0 mg / L.

7. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond according to claim 6, is characterized in that, In step S300, the water source used to dilute the compound bacterial solution is the surface water of the aquaculture pond; and in the mixture, glucose with a mass fraction of 0.2%-0.5% is added as an inorganic salt osmotic pressure regulator.

8. The process for improving the low mortality rate of aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond according to claim 7, is characterized in that, In step S300, the specific operation of "transferring to the aquaculture pond" after immersion is as follows: After the set immersion time is reached, the original pond water of the breeding pond is gradually introduced into the immersion container containing the mixture at a flow rate of 5-10 L / min until the volume ratio of the original pond water in the mixture reaches 70%-80%. Then, the water introduction is stopped, and the mixture is left to stand for 5-10 minutes before the seedlings are released into the breeding pond with the wind.

9. The process for improving low-mortality aquaculture ponds by introducing compound microbial inoculum into the pond and removing the fish from the pond according to claim 8, is characterized in that, In step S400, the potassium persulfate compound salt used is a specially prepared bottom sedimentation slow-release tablet with an effective active oxygen mass fraction of not less than 10%, and the amount of scattering is controlled at 1.0-1.5 kg / mu, and the scattering area is mainly concentrated in the feeding area and deep water sewage discharge area of ​​the aquaculture pond. In step S400, the particle size of the zeolite powder is limited to 60-80 mesh; the specific operation process of "mixed adsorption" is as follows: the composite bacterial liquid is evenly sprayed onto the surface of the zeolite powder in the form of high pressure micro-mist, while spraying and stirring, and then left to stand and solidify in a cool, dark place for 2-3 hours. After the surface is free of water and has a loose granular shape, the bottom is then sprinkled.

10. The process for improving low-mortality aquaculture ponds by introducing compound microbial inoculum solution into the pond and removing the fish from the pond according to claim 9, is characterized in that, In step S400, the specific preparation process of the feed mixed with compound bacterial solution and immunopolysaccharide is as follows: based on the mass of the conventional feed as 100%, 2.0% of the compound bacterial solution and 0.3%-0.5% of yeast β-glucan are added sequentially as immunopolysaccharides. After thorough mixing and absorption, 1.0%-1.5% of refined fish oil is used to spray and coat the surface of the feed.

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