An integrated farming method for reducing the incidence of disease in the farming of tilapia

Abstract

This invention discloses an integrated farming method for reducing the incidence of diseases in tilapia, comprising: configuring farming facilities; pre-culturing water and colonizing microorganisms before stocking; phased lighting control; dynamic water quality control and microbial enhancement; acoustic conditioning and feeding optimization; coordinated management of recirculating water and wastewater discharge; and adding a feed additive composed of activated carbon and Bacillus licheniformis to the basic feed, wherein the activated carbon is 100-300 mesh powder and the concentration of Bacillus licheniformis is 10%. 5 -10 10 CFU / g, activated carbon is pre-mixed with sodium alginate aqueous solution and added to the feed, and bacterial solution is sprayed on top; it is prepared and used immediately. This invention systematically integrates multiple environmental stress factors and activated carbon-Bacillus licheniformis composite additives to significantly reduce the incidence of diseases and improve growth performance in tilapia farming.

Description

Technical Field

[0001] This invention relates to the field of aquaculture technology, specifically to an integrated aquaculture method for reducing the incidence of diseases in tilapia farming. Background Technology

[0002] Tilapia is an important freshwater aquaculture species in my country and globally, favored by farmers for its rapid growth, strong adaptability, and high feed conversion rate. However, with the continuous expansion of aquaculture scale and increasing stocking density in recent years, the aquatic environment has deteriorated, and feed quality has become increasingly difficult to guarantee. The combination of these adverse factors has led to a significant decrease in tilapia survival rates and made the aquaculture process increasingly challenging. Under high-density aquaculture conditions, large amounts of uneaten feed and feces accumulate, ammonia nitrogen and nitrite concentrations rise, water quality deteriorates frequently, and fish remain in a sub-healthy state for extended periods, resulting in weakened immunity and increased susceptibility to various diseases (such as streptococcal disease and parasitic diseases), causing substantial economic losses. Existing technologies cannot simultaneously address the cascading problems of water quality fluctuations, decreased feeding due to stress, intestinal flora imbalance, weakened immunity, and high incidence of streptococcal disease. This invention addresses these pain points by providing an integrated, synergistic, and practical low-disease tilapia aquaculture method.

[0003] Therefore, there is an urgent need for a more mature and stable tilapia farming method that can systematically improve the farming environment, reduce the incidence of diseases, and increase the survival rate of farmed tilapia in order to meet the current development needs of intensive aquaculture. Summary of the Invention

[0004] The purpose of this application is to overcome the shortcomings and deficiencies of the existing technology and provide an integrated farming method to reduce the incidence of diseases in tilapia farming.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solution:

[0006] An integrated farming method for reducing the incidence of diseases in tilapia farming includes the following steps:

[0007] (1) Configuration of aquaculture system: Select a soil pond or a high-level aquaculture pond with a fully plastic film bottom and an independently controllable bottom water inlet and outlet system. Set a vertical sewage outlet at the lowest point of the pond bottom. Install a three-channel programmable LED array lighting system above the pond surface. The system has independently controllable red, yellow and white LED light groups. Install a set of omnidirectional underwater moving coil loudspeakers at equal intervals on the inner side of the pond wall and 0.3-0.5m below the normal water level to form a sound wave emission array. Connect an integrated water treatment device in parallel between the inlet and outlet main pipes of the aquaculture pond. The device is equipped with a rotary microfilter, an immobilized biological carrier packing tower and a low-pressure high-output ultraviolet disinfection unit in sequence along the water flow direction. The packing tower is pre-enriched with a complex microbial community, which is composed of Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria and nitrifying bacteria.

[0008] (2) Pre-culture before stocking: 18-22 days before stocking, fill the breeding pond with water to a depth of 0.6-0.8m, start the integrated water treatment device to circulate the water, and turn on the oxygenation system for 7-10 days to complete the activation and proliferation of the complex microbial community; replenish the water source to a depth of 1.2-1.5m and adjust the pH value of the water to 7.2-8.0; select healthy tilapia fry with a body length of 5-8cm, disinfect them by soaking in light salt water with sodium chloride, and transfer them to the breeding pond at a density of 80-120 fish / ㎡;

[0009] (3) Phased light control: The first 30 days after the seedlings are introduced into the pond is the early adaptation and domestication stage. Yellow LED lights are turned on daily with a photoperiod of 14L:10D. The 31st to 90th days after the seedlings are introduced into the pond is the mid-term rapid accumulation and growth stage. Yellow and red lights are used alternately for irradiation with a photoperiod of 16L:8D. The 91st day after the seedlings are introduced into the pond to the day of harvest is the late meat quality control stage. Only yellow LED lights are turned on with a photoperiod of 12L:12D.

[0010] (4) Dynamic water quality control and microbial enhancement: The integrated water treatment device is kept running continuously throughout the entire aquaculture cycle to maintain dissolved oxygen at no less than 5.0 mg / L; when ammonia nitrogen exceeds 0.5 mg / L or nitrite exceeds 0.1 mg / L, the emergency response procedure is initiated: the circulating water flow rate of the packing tower is increased to 1.5-2.0 times the normal flow rate, exogenous carbon source substances are added to the water body to raise the carbon-nitrogen ratio to above 10, and continuous enhanced aeration is started for no less than 24 hours; pre-activated compound microbial liquid inoculant is added to the aquaculture water body every 5-8 days;

[0011] (5) Sound wave training and feeding optimization: On the 4th day after the fry are introduced into the pond, the conditioned reflex between sound waves and feeding is established. The underwater speaker array is turned on 3-5 minutes before the planned feeding time each day, and the sound wave signal of 70-100 Hz is played continuously until the end of feeding. When encountering severe weather conditions during the breeding period, the high-frequency sound wave of 110-140 Hz is played to stimulate the tilapia and enhance its adaptability to environmental stress.

[0012] (6) Coordinated management of circulating water and sewage discharge: A semi-closed aquaculture water management mode is adopted throughout the entire aquaculture cycle, with water recycling as the main method. The daily circulating water volume is not less than 200% of the total water volume of the aquaculture pond. Bottom sewage discharge is carried out once every 12-18 days in the evening after feeding. Within 24-48 hours after each sewage discharge, the circulating water flow rate of the packing tower is temporarily increased to 1.2-1.5 times the normal flow rate, and an exogenous carbon source is added once.

[0013] (7) Feeding with functional feed additives: Throughout the entire breeding cycle, tilapia-specific extruded compound feed is used as the base feed, with the addition of compound feed additives; the compound additives are composed of activated carbon and Bacillus licheniformis, the activated carbon is 100-300 mesh powder, and the concentration of Bacillus licheniformis is 10 mg / gram of feed. 5 -10 10 CFU; In the feeding cycle, feed containing compound feed additives is continuously fed for 5-7 days every 30 days, and only basic feed is fed for the rest of the time.

[0014] In one or more embodiments, the single-mouth area of ​​the aquaculture pond in step (1) is 1200-600 square meters, the pond depth is 2.0-2.5 meters, the effective water depth is 1.5-1.8 meters, and the bottom of the pond forms a gentle slope from the center to the surrounding area with a slope of 2-5°.

[0015] In one or more embodiments, the peak wavelength of the red LED light group in step (1) is 620-660nm, the peak wavelength of the yellow LED light group is 580-600nm, and the color temperature of the white LED light group is 4000-5000 K; the installation height of each light group is 1.0-1.5 m above the water surface, and the rated power density is 5-12W / m²; the switching sequence and output power ratio of the lighting system are controlled by a programmable logic controller with a preset phased lighting parameter adjustment algorithm.

[0016] In one or more embodiments, the immobilized biological carrier packing tower described in step (1) is filled with a polyurethane-based porous carrier material with a specific surface area of ​​not less than 800 m² / m³. A composite microbial community is enriched within the pores of the carrier material through a circulating biofilm method. The community is composed of Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria, and nitrifying bacteria in a viable ratio of 3:2:2:1. The adhesion density of the microbial community on the carrier surface is not less than 1 × 10¹² per gram of carrier material. 8 CFU.

[0017] In one or more embodiments, during the early adaptive acclimatization phase described in step (3), the output power of the yellow LED lamp group linearly increases to the set value within the first 20 minutes after startup and linearly decreases to zero within the first 20 minutes before shutdown during the daily lighting period.

[0018] In one or more embodiments, the exogenous carbon source in step (4) is one or more of brown sugar, molasses or sodium acetate, and the carbon source is pre-dissolved at a dose of 5-15 g / m³ and then evenly sprinkled throughout the pool.

[0019] In one or more embodiments, the compound microbial liquid inoculant in step (4) is prepared from Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria and nitrifying bacteria in a viable bacteria ratio of 3:2:2:1, with a total viable bacteria concentration of not less than 1×10⁻⁶. 8 The dosage is 10-25 ml / m³, CFU / ml; oxygenation should be suspended for 1 hour before the dosage is applied.

[0020] In one or more implementations, after the sound wave and feeding conditioned reflex are established in step (5), starting from the middle period, the sound wave signal is played only before the first two feedings each day, and not played during the third feeding.

[0021] In one or more implementation schemes, in step (6), the amount of fresh water replenished from the external water source each day accounts for 3-8% of the total water volume of the aquaculture pond, and each bottom sludge discharge lasts for 30-50 minutes until the discharged water changes from turbid to clear.

[0022] In one or more embodiments, the method of adding compound feed additives to the basic feed in step (7) is as follows: activated carbon is mixed with 1.3% to 1.8% (w / v) sodium alginate aqueous solution to form an activated carbon-sodium alginate mixture, and then added to the basic feed at a ratio of 1.5% to 3.0% and dried; the Bacillus licheniformis is resuspended in PBS buffer and then sprayed on, and the feed is prepared and used immediately.

[0023] Compared with related technologies, the present invention has the following advantages:

[0024] (1) This invention organically integrates and synergistically links five technical modules: programmed phased regulation of light rhythm, compound formulation and periodic activation addition of in-situ microbial agents, hydraulic ecological regulation of intermittent circulating water combined with timed bottom sewage discharge, conditioned reflex domestication mediated by sound wave signals, and activated carbon-Bacillus licheniformis compound feed additive. This achieves systematic control of multiple environmental stress factors during the breeding cycle and significantly reduces the incidence of diseases in tilapia throughout the breeding cycle.

[0025] (2) This invention provides a light environment that matches the physiological needs of tilapia at each stage of aquaculture by using programmed phased regulation of light rhythm—in the early stage, yellow light is used to promote feeding and the establishment of basic immunity; in the middle stage, yellow and red light are used alternately to promote digestive enzyme activity and growth rate; and in the later stage, light intensity is reduced to control fat deposition.

[0026] (3) This invention improves the feeding concentration of tilapia by using the correlation between sound waves and feeding to reduce the amount of uneaten food. At the same time, it enhances the adaptability of tilapia to environmental stresses such as sudden temperature changes and heavy rainfall by using high-frequency sound wave stress training under severe weather conditions, thereby reducing the stress response level.

[0027] (4) This invention achieves long-term stable control of ammonia nitrogen and nitrite in aquaculture water by combining in-situ purification of composite microbial communities with immobilized biological carrier packing tower, periodic replenishment of liquid bacterial agents, timed bottom sewage discharge, and on-demand carbon source replenishment.

[0028] (5) The method of the present invention reduces the frequency and total amount of use of chemical disinfectants and antibiotics, and reduces the nitrogen and phosphorus pollution load in aquaculture wastewater, which is in line with the technical development direction of green and low-carbon aquaculture.

[0029] The inventiveness of this invention lies in the systematic coupling of light environment regulation, sound wave anti-stress, aquatic microbial ecology, and activated carbon carrier probiotic slow release to achieve a triple disease prevention mechanism of "stable external environment + healthy intestinal tract + strong immunity". This is not a simple superposition and has non-obviousness.

[0030] To provide a clearer understanding of this application, the specific embodiments of this application will be described below in conjunction with the accompanying drawings. Attached Figure Description

[0031] Figure 1 This is a comparison chart of growth indicators and survival rates of tilapia after being fed different feeds in Example 2.

[0032] Figure 2 This is a comparison of muscle texture in tilapia after being fed different diets in Example 2.

[0033] Figure 3This is a comparative graph showing the effects of different feeds on the antioxidant index of tilapia serum in Example 2.

[0034] Figure 4 The images show a comparison of the effects of different feeds on the intestinal structure of tilapia in Example 2. A represents a section of intestinal tissue from the control group (H&E staining, scale bar = 20 μm); B represents a section of intestinal tissue from the BL group (H&E staining, scale bar = 50 μm); C represents a section of intestinal tissue from the AB group (H&E staining, scale bar = 50 μm); and D represents a comparison of villus height and intestinal wall thickness in tilapia after feeding different feeds. VH represents villus height, and MT represents intestinal wall thickness.

[0035] Figure 5 The diagram shows the comparative effects of different feeds on the structure of the gut microbiota in tilapia in Example 2. In this diagram, A is a PCoA scatter plot based on the Beta diversity index between samples, and B is a diagram of gut microbiota composition at the genus level.

[0036] Figure 6 The images show the effects of different feeds on the expression of immune-related genes in tilapia in Example 2. A represents a comparison of quantitative fluorescence expression of immune genes in tilapia liver tissue; B represents a comparison of quantitative fluorescence expression of immune genes in tilapia spleen tissue; and C represents a comparison of quantitative fluorescence expression of immune genes in tilapia intestinal tissue.

[0037] Figure 7 This is a comparison diagram showing the effects of different diets on the quantitative expression of liver lipid metabolism genes in tilapia in Example 2.

[0038] Figure 8 This is a comparison chart showing the relative survival rate (RPS) of tilapia after being challenged with different feeds in Example 2. Detailed Implementation

[0039] To make the objectives, technical solutions, and advantages of this application clearer, the embodiments of this application will be described in further detail below with reference to the accompanying drawings.

[0040] It should be understood that the described embodiments are merely some, not all, of the embodiments of this application. All other embodiments obtained by those skilled in the art based on the embodiments of this application without creative effort are within the scope of protection of the embodiments of this application.

[0041] In the following description, when referring to the accompanying drawings, unless otherwise indicated, the same numbers in different drawings represent the same or similar elements. In the description of this application, it should be understood that the terms "first," "second," "third," etc., are used only to distinguish similar objects and are not necessarily used to describe a specific order or sequence, nor should they be construed as indicating or implying relative importance. Those skilled in the art can understand the specific meaning of the above terms in this application according to the specific circumstances. The singular forms "a," "the," and "the" used in this application and the appended claims are also intended to include the plural forms, unless the context clearly indicates otherwise. The word "if" as used herein can be interpreted as "when," "when," or "in response to determination."

[0042] Furthermore, in the description of this application, unless otherwise stated, "multiple" means two or more. "And / or" describes the relationship between related objects, indicating that three relationships can exist. For example, A and / or B can represent: A alone, A and B simultaneously, or B alone. The character " / " generally indicates that the preceding and following related objects have an "or" relationship.

[0043] The tilapia species used in the following examples, comparative examples, and test examples are all Nile tilapia.

[0044] In Example 2 below, statistical analysis was performed using GraphPad Prism 10. One-way ANOVA and Duncan's multiple comparisons were used for comparisons between groups. The sample size for each indicator was n=3, and the significance level was set at α=0.05. Statistics are expressed as mean ± standard deviation (mean ± SD).

[0045] Example 1

[0046] An integrated farming method for reducing the incidence of diseases in tilapia farming includes the following steps:

[0047] (1) Configure the aquaculture system

[0048] Elevated aquaculture ponds with a fully plastic-film bottom were selected. The pond bottom was hardened, and the inlet and outlet drainage systems were independently controllable. Each pond has an area of ​​2400 square meters, a depth of 2.2 meters, and an effective water depth of 1.6 meters. The pond bottom slopes gently at 3.5% from the center outwards. A vertical drain pipe is installed at the lowest point of the bottom, controlled by both an electric butterfly valve and manual assistance. A three-channel programmable LED array lighting system is installed above the pond surface, featuring independently controllable red LED light groups (peak wavelength 630nm), yellow LED light groups (peak wavelength 590nm), and white LED light groups (color temperature 4500K). The light groups are installed 1.2 meters above the water surface, with a rated power density of 8W / m². The switching sequence and output power ratio of the lighting system are controlled by a programmable logic controller with a preset phased lighting parameter adjustment algorithm. A set of omnidirectional underwater moving-coil loudspeakers is installed at equal intervals along the inner sides of the four walls of the aquaculture pond, 0.4 meters below the normal water level. The spacing between adjacent loudspeakers is 6 meters, each loudspeaker has a rated power of 20W, and its orientation is independently adjustable. All loudspeakers emit sound towards the geometric center of the aquaculture pond, forming a sound wave emission array. An integrated water treatment device is connected in parallel between the inlet and outlet mains of the aquaculture pond. This device, arranged sequentially along the water flow direction, includes a rotary microfilter, an immobilized biological carrier tower, and a low-pressure, high-output ultraviolet disinfection unit. The tower is filled with a polyurethane-based porous carrier material with a specific surface area of ​​850 m² / m³. A composite microbial community is pre-enriched within the carrier pores using a circulating biofilm method. The community consists of Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria, and nitrifying bacteria in a viable ratio of 3:2:2:1, with a bacterial density on the carrier surface of no less than 1×10⁻⁶. 8 CFU / g vector.

[0049] (2) Pre-breeding before release

[0050] Twenty days before stocking the fry, fill the rearing pond with water to a depth of 0.7 meters, activate the integrated water treatment device to circulate the water, and turn on the aeration system to maintain dissolved oxygen at no less than 5.0 mg / L for 8 days to complete the activation and proliferation of the complex microbial community. Then, add water to a depth of 1.4 meters, adjust the pH of the water to 7.5 with baking soda, and maintain the total alkalinity at no less than 80 mg / L (calcium carbonate equivalent). Select 2400 Nile tilapia fry with a body length of 6-7 cm, intact body surface, active swimming, and no lesions, disinfect them by soaking them in 0.8% sodium chloride saline solution for 10 minutes, and transfer them to the rearing pond at a density of 100 fry / m².

[0051] (3) Phased light regulation

[0052] The first 30 days after the seedlings are introduced into the pond are the initial acclimatization and domestication phase: Yellow LED lights are turned on daily at a light intensity of 260 lx, with a photoperiod of 14 L:10 D (lighting period 5:30-19:30). During the daily light period, the output power of the yellow lights linearly increases to the set value within the first 20 minutes after startup and linearly decreases to zero within the first 20 minutes before shutdown. The next 31-90 days after the seedlings are introduced into the pond are the mid-term rapid growth phase: alternating yellow and red light is used. Yellow light is turned on daily from 5:00-9:00 and 17:00-21:00 at a light intensity of 300 lx; ​​red light is turned on from 9:00-17:00 at a light intensity of 200 lx. The photoperiod is 16 L:8 D (lighting period 5:00-21:00). The period from the 91st day after the seedlings are introduced into the pond to the day they are harvested is the later stage of meat quality control: only yellow light is turned on, with a light intensity of 180 lx and a photoperiod of 12L:12D (lighting period 6:00-18:00).

[0053] (4) Dynamic water quality control and microbial enhancement

[0054] Throughout the entire aquaculture cycle, the integrated water treatment system is kept running continuously. Dissolved oxygen, pH, ammonia nitrogen, and nitrite levels in the water are measured daily using a portable multi-parameter water quality analyzer. Dissolved oxygen is maintained at no less than 5.0 mg / L through the microporous aeration network at the bottom of the pond and the water treatment system's built-in oxygenation system. Additional auxiliary oxygenation is activated daily from 5:00-7:00 AM and 6:00-8:00 PM to maintain dissolved oxygen at 6.0-7.0 mg / L. When ammonia nitrogen exceeds 0.5 mg / L or nitrite exceeds 0.1 mg / L, emergency measures are initiated: first, the circulating water flow rate in the packing tower is increased to 1.8 times the normal flow rate; then, brown sugar is added to the aquaculture water, pre-dissolving 10 g / m³ in a small amount of water and then evenly sprinkled throughout the pond to adjust the carbon-to-nitrogen ratio to above 10; finally, continuous enhanced aeration is activated through the microporous aeration network for at least 24 hours. A compound microbial liquid inoculant is added to the aquaculture water every 6 days. This microbial agent is formulated from Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria, and nitrifying bacteria in a viable bacteria ratio of 3:2:2:1, with a total viable bacteria concentration of not less than 1×10⁻⁶. 8 The dosage is 18 mL / m³, CFU / mL. The administration should be carried out at 10:00 AM on a clear day. Aeration should be suspended for 1 hour before administration and resumed immediately after administration.

[0055] (5) Sound wave training and feeding optimization

[0056] On the fourth day after tilapia fry were introduced into the pond, a conditioned reflex linking sound waves and feeding was established. Four minutes before the first three feedings each day, an underwater speaker array was activated, continuously playing a sine wave signal at 85 Hz and a sound pressure level of 90 dB until the end of feeding. During the early acclimatization and rapid growth phases, feeding was conducted three times daily (8:00, 12:30, and 17:30), with each feeding lasting no more than 25 minutes. During the later stages of meat quality control and before harvesting, feeding was conducted twice daily (8:30 and 16:30), with each feeding lasting 30 minutes. During periods of continuous rainy weather, a 120 Hz, 100 dB high-frequency sound wave stimulus was played at a fixed time each day for 12 minutes, once daily for two consecutive days.

[0057] (6) Coordinated management of circulating water and sewage discharge

[0058] Throughout the entire aquaculture cycle, a semi-closed aquaculture water management model is adopted, primarily based on water recycling and supplemented with small amounts of fresh water. The daily circulating water volume treated by the integrated water treatment device is no less than 200% of the total water volume of the aquaculture pond (i.e., at least two circulations per day), and the daily fresh water supplementation is controlled at 5% of the total water volume. Bottom sludge is discharged every 15 days, between 20:00 and 21:00 after feeding. During sludge discharge, the electric butterfly valve at the vertical sludge drain at the bottom of the pond and the submersible pump in the sludge collection well are opened and kept open for 40 minutes until the discharged water changes from turbid to clear. After sludge discharge, the butterfly valve is closed, and an equal amount of fresh water is added to the original water depth. Within 24-48 hours after each sludge discharge, the circulating water flow rate in the packing tower is temporarily increased to 1.3 times the normal flow rate, and brown sugar is added once at a rate of 10 g / m³.

[0059] (7) Feeding with functional feed additives

[0060] Throughout the entire aquaculture cycle, tilapia-specific extruded compound feed was used as the base feed, containing 30% crude protein and 5% crude fat. A compound feed additive was prepared: 1 kg of 200-mesh activated carbon powder was mixed with 2.5 kg of a 1.5% (w / v) sodium alginate aqueous solution to form an activated carbon-sodium alginate mixture. This mixture was added to 100 kg of the base feed at a ratio of 2.0%, mixed thoroughly, and dried at 35°C. Bacillus licheniformis powder was resuspended in sterile PBS to a final concentration of 10... 8 Apply CFU / mL evenly to the surface of the dried feed at a ratio of 10 mL / kg feed, turning the feed over as you go. Use immediately after preparation. Each 30-day period is considered an addition cycle, with 6 consecutive days of feeding. The remaining time, only the basic feed should be given.

[0061] Example 2

[0062] An integrated farming method for reducing the incidence of disease in tilapia farming is presented. The steps are basically the same as in Example 1, except that the preparation parameters of the compound feed additive are different in this example: 100-mesh activated carbon is used, the sodium alginate aqueous solution concentration is 1.3% (w / v), the addition ratio of the activated carbon-sodium alginate mixture in the basic feed is 1.5 wt%, and the concentration of Bacillus licheniformis is 10%. 5 CFU / g; all other parameters remain the same as in Example 1.

[0063] Comparative Example 1

[0064] This comparative example demonstrates a conventional tilapia pond culture method. A 2400 square meter pond with a depth of 2.0 meters and an effective water depth of 1.5 meters was selected. Before stocking, the pond was disinfected with quicklime, and 2400 Nile tilapia fry (6-7 cm in length) were stocked. Conventional management methods were employed during the culture period: feed was administered 2-3 times daily; aeration was provided by a paddlewheel aerator for 8-10 hours daily; 30% of the water was changed every 10 days; no light control or acoustic conditioning was used; no immobilized microbial carrier towers or compound bacterial solutions were used for water purification; and the culture cycle remained at 150 days.

[0065] Comparative Example 2

[0066] A method for tilapia farming is provided, with steps largely the same as in Example 1, except that this comparative example does not involve phased control of the light rhythm, the LED lighting system is not turned on throughout the process, only natural light is used, and no phased changes in light parameters are set. All other technical parameters remain consistent with those in Example 1.

[0067] Comparative Example 3

[0068] A method for tilapia farming is basically the same as that in Example 1, except that this comparative example does not involve sound-mediated conditioned reflex training, does not install an underwater speaker array, does not play sound signals when feeding, and does not establish a conditioned reflex association between sound waves and feeding. All other technical parameters are consistent with those in Example 1.

[0069] Comparative Example 4

[0070] A method for tilapia farming is provided, with steps largely the same as in Example 1, except that this comparative example does not involve the immobilization and periodic replenishment of the composite microbial community. The immobilized biological carrier packing tower in the integrated water treatment device does not pre-enrich the composite microbial community, and no composite microbial liquid inoculant is added to the water during the farming period. All other technical parameters remain consistent with those in Example 1.

[0071] Comparative Example 5

[0072] A method for tilapia farming is provided, with steps largely the same as in Example 1, except that this comparative example only feeds the fish with basic feed throughout the entire farming cycle, without adding activated carbon-Bacillus licheniformis complex (compound feed additive). All other technical parameters remain consistent with those in Example 1.

[0073] Comparative Example 6

[0074] A method for tilapia farming is basically the same as that in Example 1, except that activated carbon is added alone without Bacillus licheniformis. The activated carbon-sodium alginate mixture is added to the basic feed in the same proportion, dried, and then fed. Bacillus licheniformis bacterial solution is not sprayed. Other parameters are the same as in Example 1.

[0075] Comparative Example 7

[0076] A method for tilapia farming is basically the same as that in Example 1, except that Bacillus licheniformis is added alone without activated carbon. The Bacillus licheniformis is directly resuspended in PBS and sprayed onto the surface of the basic feed without adding the activated carbon-sodium alginate mixture. Other parameters are consistent with those in Example 1.

[0077] Detection Example 1

[0078] Parallel comparative experiments were conducted on the various tilapia farming methods of Examples 1-2 and Comparative Examples 1-7, with three replicates for each method and a uniform farming period of 150 days. After the farming period, the survival rate, disease incidence, and feed conversion efficiency of the tilapia in each group were statistically analyzed. The results are shown in Tables 1 and 2.

[0079] Table 1. Comparison of tilapia survival rate and disease incidence under different farming methods

[0080]

[0081] The data above show that the survival rate of tilapia using the farming methods of Examples 1 and 2 of this invention is significantly higher than that using the farming methods of Comparative Examples 1-7, indicating that the combined use of activated carbon and Bacillus licheniformis in the methods of this invention produces a synergistic effect, and neither can be omitted.

[0082] Table 2 Comparison of feed conversion efficiency under different farming methods

[0083]

[0084] The data above show that the feed conversion ratio of tilapia using the farming methods of Examples 1 and 2 of this invention is significantly higher than that of the farming methods of Comparative Examples 1-7, further verifying the superiority of the compound additive described in this invention.

[0085] The above results show that the present invention, through the synergistic effect of five technical means—staged illumination, acoustic stress regulation, circulating water treatment, microbial enhancement, and activated carbon-Bacillus licheniformis composite additive—increases the survival rate of tilapia by more than 19.2%, reduces the feed conversion ratio by more than 0.2, and reduces the disease incidence rate by more than 80% compared with conventional farming, thus achieving antibiotic-free and healthy farming. Each technical link is indispensable, demonstrating significant synergistic effects and outstanding inventiveness.

[0086] Detection Example 2

[0087] This test case examined various indicators of tilapia after being fed different feeds (56 days) to demonstrate that the activated carbon-Bacillus complex additive brought significant positive effects.

[0088] 2.1 Experimental fish and their husbandry management

[0089] Nile tilapia were fed commercial feed (Fujian Tianma Technology Group) at a rate of 1% of their body weight daily. Three hundred uniformly sized, healthy Nile tilapia were randomly selected and divided into three groups: control group (control), Bacillus licheniformis group (BL), and activated carbon-Bacillus licheniformis compound group (AB). Fish were manually fed twice daily, and any dead fish were recorded and removed.

[0090] 2.2 Experimental Feed Preparation

[0091] A 1.5% sodium alginate aqueous solution (w / v) was added to 200-mesh activated carbon at a ratio of 1.5%. After thorough mixing, a mixture of activated carbon and sodium alginate was obtained. This mixture was added to the feed at a ratio of 2%, thoroughly mixed, and dried overnight using a dryer to obtain the experimental feed for group AC.

[0092] To avoid sodium alginate becoming a variable, a 1.5% sodium alginate aqueous solution (w / v) was added to the feed at a ratio of 0.03%, water was added and mixed well, and the feed was dried overnight using a dryer to obtain the control group experimental feed.

[0093] The BL group experimental diet consisted of the same diet as the control group, but was sprayed with bacterial particles resuspended in PBS at a concentration of 10. 7 Bacillus licheniformis at CFU / g. Prepare and use immediately.

[0094] The experimental diets of groups AB, in addition to those of groups AC, were sprayed with bacterial particles resuspended in PBS at a concentration of 10. 7 Bacillus licheniformis at CFU / g. Prepare and use immediately.

[0095] 2.3 Oral administration experimental design

[0096] In the three experimental groups, the Control group, BL group and AB group were fed commercial feed in odd-numbered weeks (1, 3, 5, 7) and treated feed in even-numbered weeks (2, 4, 6, 8).

[0097] 2.4 Tissue and serum sample collection

[0098] 24 hours after the experiment, no further feeding was given. Before sampling, each group of experimental fish was weighed and counted to calculate growth performance indicators. Three fish were randomly selected from each group and anesthetized with eugenol (Shanghai Pharmaceutical Group Co., Ltd.). Blood was then collected from the tail vein and placed flat on a table for 30 minutes, followed by incubation at 4°C overnight. The next day, the samples were centrifuged at 3500 rpm for 2 minutes, and the serum was aspirated into 1.5 ml centrifuge tubes and stored at -80°C for related enzyme activity assays. Additionally, from each group, three fish were randomly selected, and the midgut was harvested, gently washed with physiological saline, and fixed with tissue fixative for intestinal structure analysis. Intact intestinal tissue was preserved in sterile tubes and frozen at -80°C for subsequent intestinal flora analysis. Simultaneously, from each group, three fish were randomly selected, and muscle tissue was harvested for muscle texture analysis. Finally, from each group, three fish were randomly selected, and their liver, spleen, and intestinal tissue were harvested and frozen at -80°C for related gene expression analysis.

[0099] 2.5 Testing and Results of Various Indicators

[0100] 2.5.1 Detection of growth indicators

[0101] Feeding was stopped the day before the measurement, and 30 fish were randomly selected for weight measurement. The formulas for calculating weight gain rate (WGR), specific growth rate (SGR), feed conversion ratio (FCR), and survival rate (SR) are as follows:

[0102]

[0103]

[0104]

[0105]

[0106] Results: The activated carbon-Bacillus complex promoted growth.

[0107] The results of a feeding experiment on Nile tilapia with different diets after 56 days, including weight gain (WGR), specific growth rate (SGR), feed conversion ratio (FCR), and survival rate (SR), are as follows: Figure 1As shown in the figure. Statistical analysis showed that the WGR and SGR of the BL and AB groups were significantly higher than those of the control group (p < 0.05), while there was no significant difference between the BL and AB groups (p > 0.05). The FCR of the BL and AB groups was significantly lower than that of the control group (p < 0.05), while there was no significant difference between the BL and AB groups (p > 0.05). There was no significant difference in SR between the control group and the BL and AB groups (p > 0.05), but the BL and AB groups were slightly higher than those of the control group.

[0108] 2.5.2 Muscle Texture Detection

[0109] Muscle samples were analyzed using a texture analyzer in TPA (texture profile analysis) mode with a P / 10 probe. The initial compression speed was 2 mm / s, the compression speed was 1 mm / s, and the compression speed was 2 mm / s after the test, with a 5-s interval between compressions. The hardness, adhesiveness, springiness, and chewiness of the muscle can be directly obtained from the TPA curve.

[0110] Results: The activated charcoal-Bacillus complex improved muscle texture.

[0111] The results of different diets on Nile tilapia after 56 days of feeding experiments, including hardness, adhesiveness, springiness, and chewiness, are as follows: Figure 2 As shown, there was no significant difference in Hardness between the control group and the BL and AB groups (p > 0.05). The AB group had the lowest absolute Adhesiveness value, which was significantly different from both the control and BL groups (p < 0.05); the BL group had a significantly higher absolute Adhesiveness value than the control group (p < 0.05). The AB group had a significantly higher Springiness value than the control group (p < 0.05), while the BL group had a significantly lower Springiness value than the control group (p < 0.05), and there was also a significant difference between the two groups (p < 0.05). Compared with the control group, the BL group had a significantly lower Chewiness value (p < 0.05), and the AB group also had a significantly lower Chewiness value than the control group (p < 0.05), but the AB group had a significantly higher Chewiness value than the BL group (p < 0.05).

[0112] 2.5.3 Serum Antioxidant Stress Capacity Analysis

[0113] In this test, serum biochemical indicators were measured using a reagent kit (Nanjing Jiancheng). The detection parameters and corresponding product numbers are shown in Table 3. The SOD kit essentially utilizes the principle of competitive inhibition to consistently generate superoxide hydrogen ions (O2) through the xanthine-XOD system. .- A portion of the SOD in the sample is cleared, while the remainder reduces WST-1 to form colored substances. Therefore, the degree of color development is negatively correlated with SOD activity, thus achieving indirect quantification of SOD activity. The CAT kit relies on the colorimetric reaction of ammonium molybdate with hydrogen peroxide (H2O2) to indirectly measure CAT activity. The MDA kit is based on the colorimetric reaction of malondialdehyde (MDA), the end product of lipid peroxidation, with thiobarbituric acid (TBA). The T-AOC kit is based on the principle of ABTS [2,2'-azido-bis(3-ethylbenzothiazoline-6-sulfonic acid)] reacting with an oxidizing agent to form cationic free radicals. All indicators were measured according to the kit instructions, and the experiments were strictly conducted in accordance with the operating procedures to ensure the accuracy and reproducibility of the data.

[0114] Table 3. Names and catalog numbers of the reagent kits used for the tests.

[0115]

[0116] Results: The activated charcoal-Bacillus complex can promote serum antioxidant stress capacity.

[0117] Different feed treatment groups showed effects on the antioxidant enzyme activity of Nile tilapia, such as Figure 3 As shown, compared with the control group and the BL group, the AB group significantly increased SOD, CAT, and T-AOC in Nile tilapia (p < 0.05). However, there were no significant differences in SOD, CAT, and T-AOC between the control group and the BL group (p > 0.05), although the BL group was slightly higher than the control group. Regarding lipid peroxidation indicators, although the MDA content of Nile tilapia in each experimental group showed a certain decreasing trend (control > BL > AB), the differences between the groups were not significant (p > 0.05).

[0118] 2.5.4 Intestinal Structure Analysis

[0119] Intestinal samples were fixed in 4% formaldehyde fixative for 36 hours, dehydrated with fractionated ethanol at concentrations of 70%, 80%, and 100%, and then embedded in paraffin. Samples were cut into 4 μm sections using a microtome, stained with hematoxylin and eosin (H&E), and their morphology was observed using a microscopic imaging system. The villus length and muscle layer thickness of all sections were measured using ImageJ software.

[0120] Results: The activated charcoal-Bacillus complex improved the intestinal structure of Nile tilapia.

[0121] After the 56-day feeding experiment, the intestinal tissue structure slices of Nile tilapia in each group are shown below. Figure 4 As shown in A, 4B, and 4C, the results of intestinal villus height and intestinal wall thickness in Nile tilapia from groups AB, BL, and the control group are compared as follows. Figure 3 As shown in .5D, compared with the control group, the intestinal villus height was significantly increased in both the BL and AB groups (p < 0.05), and the AB group was significantly higher than the BL group (p < 0.05). Compared with the control group, the muscle layer thickness was significantly increased in the AB group (p < 0.05), while there was no significant difference between the BL group and the control and AB groups (p > 0.05).

[0122] 2.5.5 High-throughput sequencing of 16S RNA from the gut microbiota

[0123] In this study, the gut microbiota detection was performed by Guangzhou Gediao Biotechnology Co., Ltd. Genomic DNA was extracted from Nile tilapia gut samples, and the V3 and V4 regions of 16S rDNA were amplified using barcode-specific primers. The primer sequences were: (341F): CCTACGGGNGGCWGCAG; (806r): GGACTACHVGGGTATCTAAT. The PCR amplification products were gel-cleaved and quantified using a QuantiFluor™ fluorometer. The purified amplification products were mixed in equal volumes, ligated with sequencing adapters, and used to construct sequencing libraries for sequencing on an Illumina PE250. After obtaining raw reads, low-quality reads were first filtered, then assembled, and paired-end reads were spliced ​​into tags. These tags were then filtered again, resulting in clean tags. Next, clustering was performed based on the clean tags to remove chimeric tags detected during cluster alignment, ultimately yielding effective tags. After obtaining ZOTUs, ZOTU abundance statistics were performed based on effective tags. Based on the OTU sequences, analyses were conducted on abundance data, species annotation, species composition, and function. If valid groupings existed, inter-group differences were compared and statistically tested.

[0124] Results: The activated charcoal-Bacillus complex can improve the structure of the gut microbiota.

[0125] After the 56-day feeding experiment, based on the results of weighted unifrac, unweighted unifrac, Jaccard, and Bray data among the samples, by Figure 5As shown in Figure A, the first principal coordinate system (PCoA1) and the second principal coordinate system (PCoA2) explained 62.48% and 19.47% of the total variance, respectively, with a cumulative contribution of 81.95%. The control group samples were more concentrated along the positive direction of the PCoA1 axis, while the BL and AB group samples shifted towards the negative direction of PCoA1, showing a clear separation from the control group. The distribution of the BL and AB groups in the PCoA space also differed, indicating that different additive treatments significantly affected the gut microbiota structure, and the microbial composition of the BL and AB groups differed significantly from that of the control group.

[0126] In addition, such as Figure 5 As shown in B, in the control group, the relative abundance of genera such as *Cetobacterium* and *Akkermansia* was relatively high. In the BL group, the relative abundance of genera such as *Akkermansia* was lower than that in the control and AB groups. In the AB groups, the relative abundance of beneficial bacteria genera such as *Cetobacterium* and *Akkermansia* was significantly increased, while the abundance of *Bacteroides* was significantly decreased.

[0127] 2.5.6 Expression of immune-related genes in different tissues

[0128] RNA extraction and RT-qPCR detection were employed. Specifically, total RNA was extracted from liver, spleen, and intestinal tissues using RNAiso-Easy (TAKARA) according to the manufacturer's instructions. Genomic DNA was removed from the samples using the PrimeScript™ RT reagent Kit with gDNA Eraser (Perfect Real Time) (TAKARA) according to the manufacturer's instructions, and the RNA was reverse transcribed into cDNA. The cDNA obtained from reverse transcription was diluted and then used for RT-qPCR analysis. Reference sequences were obtained from the NCBI database (https: / / www.ncbi.nlm.nih.gov / ). Primers were designed using Primer Premier 6 software, and primers for real-time quantification were synthesized by Sangon Biotech (Shanghai) Co., Ltd. (https: / / www.sangon.com / ). Detailed primer sequences are provided in Table 4. The reverse transcription real-time quantitative polymerase chain reaction (RT-qPCR) used in this experiment was performed using the SYBR Green dye method on a real-time fluorescence quantitative PCR instrument, using 2... -△△CT The method processes and calculates RT-qPCR data.

[0129] Table 4 Primer Sequences for Real-Time Quantitative PCR

[0130]

[0131] β-actin = actin β; IgM = immunoglobulin M; IL-1β = interleukin 1β; IL-10 = interleukin 10; TNF-α = tumor necrosis factor α; TGF-β = transforming growth factor-β; ef1α = elongation factor 1α; ACC = acetyl-CoA carboxylase; PPARα = peroxisome proliferator-activated receptor; CPT1A = carnitine palmitoyltransferase 1A; FAS = fatty acid synthase; SREBP = sterol regulatory element-binding protein.

[0132] Results: The activated charcoal-Bacillus complex can enhance the expression of immune-related genes and lipid metabolism-related genes.

[0133] After a 56-day feeding trial, the effects of adding Bacillus licheniformis and its complex with activated charcoal to the feed on the expression of immune-related genes IgM, IL-1β, IL-10, TNF-α, and TGF-β in the liver of Nile tilapia were as follows: Figure 6 As shown in Figure A, compared with the control group, the BL group significantly upregulated the gene expression of IgM, IL-10, and TNF-α (p < 0.05) and significantly downregulated the expression of IL-1β and TGF-β (p < 0.05). The AB group significantly upregulated IL-10 while greatly inhibiting the expression of IL-1β and TGF-β (p < 0.05), with TGF-β expression being significantly lower than that in the BL group (p < 0.05). There was no significant difference in the expression levels of IgM and TNF-α between the AB group and the control group (p > 0.05).

[0134] Expression of immune-related genes in the spleen of Nile tilapia, such as Figure 6 As shown in B, the gene expression trends in the spleen were highly similar to those in the liver. The BL group significantly upregulated the expression of IgM, IL-10, and TNF-α, while significantly downregulating IL-1β and TGF-β (p < 0.05). In contrast, the AB group, apart from significantly downregulating IL-1β and TGF-β, showed no significant difference in IgM and TNF-α expression levels compared to the control group (p > 0.05). Furthermore, the IL-10 expression level in the spleen of the AB group was between that of the control and BL groups, with no significant difference (p > 0.05).

[0135] Expression of immune-related genes in the intestine of Nile tilapia, such as Figure 6As shown in Figure C, the expression patterns of intestinal mucosal immune genes are distinctly different from those in the hepatocellular carcinoma, pancreas, and spleen. In the BL group, the expression levels of all detected immune genes (IgM, IL-1β, IL-10, TNF-α, TGF-β) were not significantly different from those in the control group (p > 0.05). Conversely, the AB group significantly upregulated the expression levels of IgM, IL-10, and TNF-α (p < 0.05), while there were no significant differences in the expression levels of IL-1β and TGF-β among the groups (p > 0.05).

[0136] The addition of Bacillus licheniformis and its complex with activated carbon to the feed had a differential effect on the expression of lipid metabolism-related genes ACC, PPARα, CPT1A, FAS, and SREBP in the liver of Nile tilapia. Figure 7 As shown in the figure. Regarding ACC expression, there were no significant differences among the three groups (p > 0.05). There was no significant difference in PPARα expression between the control group and the BL group (p > 0.05), but the control group showed a significantly higher expression than the AB group (p < 0.05), while there was no significant difference between the BL group and the AB group (p > 0.05). CPT1A expression in the control group was significantly higher than in the BL group and the AB group (p < 0.05), while there was no significant difference between the latter two (p > 0.05). FAS expression in the AB group was significantly higher than in the BL group (p < 0.05), but there were no significant differences between the control group and the BL group, or between the control group and the AB group (p > 0.05). SREBP expression in the BL group was significantly higher than in the control group and the AB group (p < 0.05), while there was no significant difference between the latter two (p > 0.05).

[0137] 2.5.7 Virus Challenge Experiment

[0138] After a 56-day feeding experiment, 50 fish in each group were injected intraperitoneally with 0.1 mL of a solution containing 1×10⁻⁶ g of fish. 7 Fish were challenged with a CFU / mL Streptococcus agalactiae suspension. Mortality was recorded daily for each group after challenge until the fish in each group stabilized. Relative survival rates were calculated.

[0139] Results: The activated charcoal-Bacillus complex enhanced the resistance of tilapia to Streptococcus agalactiae infection.

[0140] After a 56-day rearing trial, Nile tilapia were challenged with Streptococcus agalactiae. The RPS (reactive protein levels) of each group of Nile tilapia were as follows: Figure 8As shown in the figure, on day 14 post-challenge, the RPS was 10.94% in the BL group and 19.22% in the AB group. Throughout the entire challenge observation period, the survival rates of the BL and AB groups were consistently higher than those of the control group. No deaths occurred in the AB group within 8-14 days, in the BL group within 9-14 days, and in the control group within 10-14 days. Comparatively, the AB group stabilized earliest.

[0141] In summary, adding activated charcoal to feed significantly promotes the growth performance of Nile tilapia, improves feed utilization efficiency and survival rate, and enhances muscle texture and serum antioxidant capacity. Activated charcoal also optimizes the intestinal microbiota structure and promotes improved intestinal tissue morphology. In terms of immune regulation, activated charcoal effectively enhances the body's specific immune response; in terms of hepatic lipid metabolism, it exhibits metabolic reprogramming characteristics of promoting lipid synthesis and inhibiting fatty acid oxidation. Challenge experiments have confirmed that activated charcoal shows good application potential in improving the disease resistance of aquatic animals. Adding a complex of activated charcoal and Bacillus licheniformis to feed significantly improves the growth performance of Nile tilapia, increases feed utilization efficiency, survival rate, muscle texture, and serum antioxidant capacity, while optimizing the intestinal microbiota structure and intestinal tissue morphology. In terms of immune regulation, this complex regulates the immune response in a tissue-specific manner, maintaining a pro-inflammatory-anti-inflammatory balance in systemic immune organs while enhancing intestinal mucosal immune defense. Furthermore, hepatic lipid metabolism exhibits adaptive reprogramming characteristics of upregulated fatty acid synthesis and downregulated oxidation. The challenge test further confirmed that there is a synergistic effect between activated carbon and Bacillus licheniformis, which can improve the disease resistance of fish.

[0142] Therefore, in the aquaculture method described in this invention, activated carbon and Bacillus licheniformis can enhance the disease resistance of fish, potentially exhibiting a synergistic effect. As a compound additive, it not only promotes fish growth, improves intestinal and muscle structure, and enhances resistance to bacterial diseases, but its core significance in aquaculture lies in expanding the application scope of activated carbon, especially for other probiotics or nutritional additives. Developing activated carbon compound additives with even higher synergistic efficiency will benefit aquaculture, promote sustainability, reduce antibiotic use, and foster the healthy development of the industry.

[0143] It should also be noted that the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0144] The above are merely embodiments of this application and are not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. An integrated aquaculture method for reducing the incidence of diseases in tilapia farming, characterized in that, Includes the following steps: (1) Configuration of aquaculture system: Select a soil pond or a high-level aquaculture pond with a fully plastic film bottom and an independently controllable bottom water inlet and outlet system. Set a vertical sewage outlet at the lowest point of the pond bottom. Install a three-channel programmable LED array lighting system above the pond surface. The system has independently controllable red, yellow and white LED light groups. Install a set of omnidirectional underwater moving coil loudspeakers at equal intervals on the inner side of the pond wall and 0.3-0.5m below the normal water level to form a sound wave emission array. Connect an integrated water treatment device in parallel between the inlet and outlet main pipes of the aquaculture pond. The device is equipped with a rotary microfilter, an immobilized biological carrier packing tower and a low-pressure high-output ultraviolet disinfection unit in sequence along the water flow direction. The packing tower is pre-enriched with a complex microbial community, which is composed of Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria and nitrifying bacteria. (2) Pre-culture before stocking: 18-22 days before stocking, fill the breeding pond with water to a depth of 0.6-0.8m, start the integrated water treatment device to circulate the water, and turn on the oxygenation system for 7-10 days to complete the activation and proliferation of the complex microbial community; replenish the water source to a depth of 1.2-1.5m and adjust the pH value of the water to 7.2-8.0; select healthy tilapia fry with a body length of 5-8cm, disinfect them by soaking in light salt water with sodium chloride, and transfer them to the breeding pond at a density of 80-120 fish / ㎡; (3) Phased light control: The first 30 days after the seedlings are introduced into the pond is the early adaptation and domestication stage. Yellow LED lights are turned on daily with a photoperiod of 14L:10D. The 31st to 90th days after the seedlings are introduced into the pond is the mid-term rapid accumulation and growth stage. Yellow and red lights are used alternately for irradiation with a photoperiod of 16L:8D. The 91st day after the seedlings are introduced into the pond to the day of harvest is the late meat quality control stage. Only yellow LED lights are turned on with a photoperiod of 12L:12D. (4) Dynamic water quality control and microbial enhancement: The integrated water treatment device is kept running continuously throughout the entire aquaculture cycle to maintain dissolved oxygen at no less than 5.0 mg / L; when ammonia nitrogen exceeds 0.5 mg / L or nitrite exceeds 0.1 mg / L, the emergency response procedure is initiated: the circulating water flow rate of the packing tower is increased to 1.5-2.0 times the normal flow rate, exogenous carbon source substances are added to the water body to raise the carbon-nitrogen ratio to above 10, and continuous enhanced aeration is started for no less than 24 hours; pre-activated compound microbial liquid inoculant is added to the aquaculture water body every 5-8 days; (5) Sound wave training and feeding optimization: On the 4th day after the fry are introduced into the pond, the conditioned reflex between sound waves and feeding is established. The underwater speaker array is turned on 3-5 minutes before the planned feeding time each day, and the sound wave signal of 70-100 Hz is played continuously until the end of feeding. When encountering severe weather conditions during the breeding period, the high-frequency sound wave of 110-140 Hz is played to stimulate the tilapia and enhance its adaptability to environmental stress. (6) Coordinated management of circulating water and sewage discharge: A semi-closed aquaculture water management mode is adopted throughout the entire aquaculture cycle, with water recycling as the main method. The daily circulating water volume is not less than 200% of the total water volume of the aquaculture pond. Bottom sewage discharge is carried out once every 12-18 days in the evening after feeding. Within 24-48 hours after each sewage discharge, the circulating water flow rate of the packing tower is temporarily increased to 1.2-1.5 times the normal flow rate, and an exogenous carbon source is added once. (7) Functional feed additive feeding: throughout the breeding cycle, the special puffed compound feed for tilapia is used as the basic feed, and a compound feed additive is added; the compound additive is composed of activated carbon and bacillus licheniformis; the activated carbon is 100-300 mesh powder, and the added concentration of bacillus licheniformis is 10 5 -10 10 CFU per gram of feed; in the feed feeding, every 30 days is a cycle, and the feed containing the compound feed additive is continuously fed for 5-7 days, and the basic feed is only fed for the rest of the time.

2. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The area of ​​a single breeding pond in step (1) is 1200-600 square meters, the pond depth is 2.0-2.5 meters, the effective water depth is 1.5-1.8 meters, and the bottom of the pond forms a gentle slope from the center to the surrounding area with a slope of 2-5°.

3. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The peak wavelength of the red LED light group in step (1) is 620-660 nm, the peak wavelength of the yellow LED light group is 580-600 nm, and the color temperature of the white LED light group is 4000-5000 K; the installation height of each light group is 1.0-1.5 m above the water surface, and the rated power density is 5-12 W / m²; the switching sequence and output power ratio of the lighting system are controlled by a programmable logic controller with a preset phased lighting parameter adjustment algorithm.

4. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The polyurethane-based porous carrier material with specific surface area not less than 800 m2 / m3 in the fixed biological carrier packing tower in step (1) is enriched with a complex microbial flora by circulating biofilm method. The flora is compounded by Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria and nitrifying bacteria in the proportion of 3:2:2:1 in terms of the number of viable bacteria. The adhesion density of the flora on the carrier surface is not less than 1×10 8 CFU per gram of carrier material.

5. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, In the early adaptive domestication stage described in step (3), the output power of the yellow LED lamp group linearly increases to the set value within the first 20 minutes after startup during the daily light period, and linearly decreases to zero within the first 20 minutes before shutdown.

6. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The exogenous carbon source mentioned in step (4) is one or more of brown sugar, molasses or sodium acetate. The carbon source is pre-dissolved at a dose of 5-15 g / m³ and then evenly sprinkled throughout the pool.

7. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The compound microbial liquid inoculant mentioned in step (4) is prepared by Bacillus subtilis, Bacillus licheniformis, photosynthetic bacteria and nitrifying bacteria in a viable bacteria ratio of 3:2:2:1, with a total viable bacteria concentration of not less than 1×10⁻⁶. 8 The dosage is 10-25 ml / m³, CFU / ml; oxygenation should be suspended for 1 hour before the dosage is applied.

8. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, After the sound wave and feeding conditional reflex are established as described in step (5), starting from the middle period, the sound wave signal is played only before the first two feedings each day, and not played during the third feeding.

9. The integrated aquaculture method for reducing the incidence of diseases in tilapia farming according to claim 1, characterized in that, In step (6), the amount of fresh water replenished from external water sources each day accounts for 3-8% of the total water volume of the aquaculture pond. Each bottom sludge discharge lasts for 30-50 minutes until the discharged water changes from turbid to clear.

10. The integrated aquaculture method for reducing the incidence of disease in tilapia farming according to claim 1, characterized in that, The method for adding compound feed additives to the basic feed in step (7) is as follows: activated carbon is mixed with 1.3% to 1.8% (w / v) sodium alginate aqueous solution to form an activated carbon-sodium alginate mixture, and then added to the basic feed at a ratio of 1.5% to 3.0% and dried; the Bacillus licheniformis is resuspended in PBS buffer and then sprayed on, and the feed is prepared and used immediately.

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