Treatment method for sleep disease in fancy carp culture

Abstract

The invention relates to the technical field of aquaculture, and discloses a treatment method for sleep diseases in fancy carp culture. According to the invention, early-stage rapid intervention is taken as a core, and in combination with the application of the improved suspension and the synergistic effect of the slow-release immune bait, the drug targeting is improved, the stress injury to fish bodies is reduced, the host immunity is enhanced, and the virus diffusion is effectively inhibited; according to the dynamic environment regulation and control technology, through real-time monitoring of water quality parameters and linkage adjustment of drug concentration and oxygenation strategies, environmental adaptability in the treatment process is ensured, drug abuse and residual pollution are reduced, and ecological balance of a water body is maintained; the cross infection of viruses is blocked by a whole-course partition management strategy, and the degradation of drug metabolites is accelerated and the treatment cycle is shortened in combination with the ecological restoration effect of the complex probiotics and the citric acid buffer agent. In addition, a treatment file database is established, the scheme is continuously improved through historical case analysis, the cure rate and operation efficiency are improved, and the comprehensive cost is reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of aquaculture, and specifically to a treatment method for sleep disease in koi breeding. Background Art

[0002] Koi sleep disease (also known as edema disease) is a linear double-stranded DNA virus, which is essentially a genetic defect. As long as it is the koi species, there is a possibility of getting the disease; its onset is rapid, and once it occurs, the mortality rate is nearly 100%; during the koi breeding process, if such problems are encountered, the koi are usually placed in a smaller cement pond, and 7‰ salt is added to the water to balance the osmotic pressure, and then some fish are left by means of group immunity. Even though the survival rate of the treatment by group immunity in the past was not high, many small-scale koi farms have imperfect facilities and do not have small cement temporary ponds; once the koi in the pond get sick, because there is no treatment plan for the earthen pond, basically all of them will die, and the economic losses caused in the industry every year are immeasurable. Therefore, a treatment method for sleep disease in koi breeding is proposed. Summary of the Invention

[0003] Aiming at the deficiencies of the prior art, the present invention provides a treatment method for sleep disease in koi breeding to solve the problems in the background art.

[0004] To achieve the above object, the present invention provides the following technical solution: A treatment method for sleep disease in koi breeding, including the following steps:

[0005] Step 1: Early rapid intervention: After ≥3% of the individuals in the koi group show symptoms of floating head, lying on the side or body surface congestion, evenly sprinkle a modified potassium monopersulfate compound salt suspension into the breeding water body within 2 hours, with an initial concentration of 0.07 ppm, and apply it at a dose of 50 grams per cubic meter of water body;

[0006] Step 2: Concentration adjustment in stages:

[0007] The first stage (0 - 24 hours after the onset): Maintain a concentration of 0.07 ppm and apply the medicine 2 times a day;

[0008] The second stage (24 - 72 hours): Increase the concentration to 0.09 ppm, apply the medicine 1 time a day, and combine with an oxygenation device to maintain the dissolved oxygen content ≥6 mg / L;

[0009] The third stage (after 72 hours): Reduce the concentration to 0.05 ppm, and simultaneously feed immune-enhancing bait containing 0.5% vitamin C once a day for 5 days;

[0010] Step 3: Dynamic environmental regulation:

[0011] Real-time monitor the pH value and temperature of the water body. When pH > 8.0, additionally add citric acid buffer (0.1 g / m 3 ) to stabilize the drug activity;

[0012] When the water temperature is lower than 18 °C, extend the administration interval of the modified potassium monopersulfate compound salt suspension to once every 36 hours;

[0013] Step 4: Determine the termination of treatment: When there are no new dead individuals within 48 consecutive hours and the food intake of koi carp resumes to more than 80% of that before the onset of the disease, stop administering the drug.

[0014] Preferably, the preparation method of the modified potassium monopersulfate compound salt suspension is as follows:

[0015] Mix AR grade potassium monopersulfate compound salt and food grade diatomaceous earth in a mass ratio of 1:0.2, and add 0.05% polyethylene glycol (PEG - 4000) as a dispersant. Treat with a high - speed shear emulsifier for 10 minutes to form a uniform suspension with a particle size ≤ 20 μm.

[0016] Preferably, the usage strategy of the aeration equipment is as follows:

[0017] During the drug administration period in the second stage (24 - 72 hours), start the aeration equipment daily after drug administration to maintain the dissolved oxygen content ≥ 6 mg / L, and the aeration duration is positively correlated with the drug concentration. Specifically:

[0018] When the concentration is 0.07 ppm, the aeration duration ≥ 4 hours / day;

[0019] When the concentration is 0.09 ppm, the aeration duration ≥ 6 hours / day.

[0020] Preferably, the preparation method of the immune - enhancing bait is as follows:

[0021] Step A, preparation of the base material: Using fish meal as the basic raw material, add 0.5% vitamin C, 1.2% β - glucan, and 0.3% allicin, and use a twin - screw extruder to make slow - release granules with a porous structure at 120 - 130 °C and a pressure of 30 - 40 MPa. The particle size of the slow - release granules is 2 - 3 mm;

[0022] Step B, coating with a composite film: Immerse the slow - release granules in a sodium alginate - chitosan composite solution (the mass ratio of sodium alginate to chitosan is 1:1, and the solvent is 2% acetic acid solution). Take them out after impregnation for 5 minutes and place them in a 3% calcium chloride solution for cross - linking and curing for 10 minutes to form a composite film with a thickness of 50 - 80 μm;

[0023] Step C, drying treatment: Hot - air dry the coated granules at 40 °C until the water content ≤ 8%.

[0024] Preferably, the citric acid buffer is added in the following manner:

[0025] Citric acid and sodium bicarbonate were premixed in a molar ratio of 1:1.5 and packaged in a biodegradable slow-release bag, which was hung at the water inlet of the aquaculture pond. The slow-release rate was controlled at 0.01 g / h·m 3 .

[0026] Preferably, in step 3, when the ammonia nitrogen concentration in the water body is ≥0.5 mg / L, a composite probiotic preparation (containing Bacillus subtilis and nitrifying bacteria, with a viable count of ≥1×10 8 CFU / g), the dose is 1g / m 3 , once a day, until the ammonia nitrogen concentration is less than 0.2 mg / L.

[0027] Preferably, the treatment termination determination further includes:

[0028] Mucus samples were collected from the surface of koi carp and tested using the test paper method. The negative rate of viral antigen was ≥95%, and there was no visible congestion or ulceration in the gill filament tissue.

[0029] Preferably, zoning and isolation management is adopted throughout the treatment process:

[0030] The diseased pond was divided into a core area (where diseased fish were concentrated) and a buffer area (where healthy fish were active). The pesticide concentration in the core area was increased by 10%, while the basic concentration was maintained in the buffer area. A physical isolation net (mesh size ≤ 1 cm) was set between the two areas.

[0031] Preferably, a treatment record database is established:

[0032] The initial symptom time, application parameters, water quality changes and cure rate data of each batch of treatment were recorded, and the concentration gradient was optimized through statistical analysis to increase the cure rate from 90% to 93%-95%.

[0033] Compared with the prior art, the present invention has the following beneficial effects:

[0034] The present invention takes early rapid intervention as the core, combines the synergistic effect of the administration of the modified suspension and the slow-release immune bait, improves the drug targeting, reduces stress damage to the fish body, and enhances the host immunity, effectively inhibiting the spread of the virus; the dynamic environmental control technology monitors water quality parameters in real time, adjusts the drug concentration and oxygenation strategy in linkage, ensures the environmental adaptability of the treatment process, reduces drug abuse and residual pollution, and maintains the ecological balance of the water body; the whole zoning management strategy blocks viral cross-infection, combines the ecological restoration effect of compound probiotics and citric acid buffer, accelerates the degradation of drug metabolites, and shortens the treatment cycle. In addition, the establishment of a treatment archive database continuously improves the plan through historical case analysis, improves the cure rate and operating efficiency, and reduces the overall cost.

[0035] Other features and advantages of the present invention will be set forth in the following description, and in part will be obvious from the description, or may be learned by practice of the present invention. The objectives and other advantages of the present invention can be realized and attained by the structure particularly pointed out in the specification, claims as well as the drawings. Description of the Drawings

[0036] Figure 1 It is a flowchart of the treatment method for sleeping disease in koi breeding according to the present invention;

[0037] Figure 2 It is a flowchart for preparing the immune-enhancing bait according to the present invention. Detailed Embodiments

[0038] The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present invention. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art in the technical field based on the embodiments of the present invention without creative efforts shall fall within the protection scope of the present invention.

[0039] Please refer to Figure 1 and Figure 2 , a treatment method for sleeping disease in koi breeding according to the present invention, realizes efficient and low-residue disease control through early rapid intervention, staged adjustment of drug concentration, linkage control of environmental parameters, and accurate determination of treatment termination; specifically includes the following contents:

[0040] I. Raw material preparation

[0041] 1. Preparation of the modified suspension

[0042] Raw material preparation and pretreatment:

[0043] Main drug: Potassium monopersulfate compound of AR grade (active oxygen content ≥ 4.5%, purity ≥ 98%), which needs to be stored away from light and passed through a 100-mesh sieve to remove lumps before use.

[0044] Auxiliary materials:

[0045] Food-grade diatomaceous earth: particle size ≤ 100μm, specific surface area ≥ 20m 2 / g, used to adsorb drug particles and improve suspension stability;

[0046] Polyethylene glycol (PEG-4000): as a dispersant to reduce the surface tension of particles and prevent agglomeration.

[0047] Mixing process:

[0048] Ratio: The main drug and diatomaceous earth are accurately weighed according to a mass ratio of 1:0.2 (for example, 1 kg of the main drug + 200 g of diatomaceous earth);

[0049] Premixing: Place the two substances in a V-type mixer and mix at a speed of 20 rpm for 15 minutes to ensure preliminary dispersion;

[0050] Dispersant addition: Add 0.05% PEG-4000 to the mixture (i.e., 0.5 g per 1 kg of the mixture), and continue mixing for 10 minutes.

[0051] Emulsification treatment and particle size control:

[0052] Equipment: High-speed shear emulsifier (model FLUKO FS-400, power 5.5 kW, adjustable speed 0 - 15000 rpm).

[0053] Operating parameters:

[0054] Rotation speed: 12000 rpm, shear head diameter 50 mm;

[0055] Treatment time: 10 minutes, carried out in two times (5 minutes each time, with a 2-minute interval for cooling).

[0056] Particle size detection:

[0057] Instrument: Laser particle size analyzer (Malvern mastersizer 3000);

[0058] Standard: The particle size (D90) of 90% of the particles in the suspension ≤ 20 μm, and the polydispersity index (PDI) < 0.3.

[0059] Verification of suspension stability:

[0060] Static experiment: Take 500 mL of the suspension and place it in a transparent glass graduated cylinder. Observe the layering situation after standing for 24 hours; the qualified standard is that the bottom sediment volume ≤ 5%;

[0061] Active oxygen retention rate: Detected by the iodometric method, used immediately after preparation, and the active oxygen loss rate ≤ 2%.

[0062] Storage and usage specifications:

[0063] Storage conditions: Store in a sealed and light-proof manner in an environment below 25°C;

[0064] Treatment before use: Stir for 5 minutes to ensure uniform dispersion.

[0065] The preparation of the modified potassium monopersulfate compound salt suspension aims to solve the problems of slow dissolution rate and uneven dispersion of traditional drugs. By optimizing the drug particle size and dispersion stability, it improves the drug utilization rate, reduces the stress response of koi caused by excessive local concentration, and at the same time ensures the effective release of active oxygen.

[0066] 2. Preparation of immune-enhanced bait

[0067] Step A, Base Material Preparation

[0068] Raw Material Ratio:

[0069] Fish meal (crude protein ≥ 60%, fat ≤ 8%) accounts for 97%.

[0070] Functional Additives:

[0071] Vitamin C (ascorbic acid, purity ≥ 99%) 0.5%;

[0072] β-glucan (yeast extract, purity ≥ 70%) 1.2%;

[0073] Allicin (diallyl trisulfide, purity ≥ 20%) 0.3%.

[0074] Mixing Process:

[0075] Use a twin-shaft paddle mixer (model HLSH-2) to mix the above raw materials and functional additives, with a rotation speed of 45 rpm, a mixing time of 20 minutes, and a coefficient of variation (CV) of mixing uniformity ≤ 5%.

[0076] Extrusion Molding:

[0077] Equipment: Use a co-rotating twin-screw extruder (model TSL-65, screw diameter 65 mm, length-diameter ratio 10:1) to extrude the mixed raw materials into slow-release granules with a porous structure.

[0078] Process Parameters:

[0079] Temperatures of the twin-screw extruder: Zone 1 120 °C, Zone 2 125 °C, Zone 3 130 °C;

[0080] Pressure of the twin-screw extruder: 30 - 40 mPa;

[0081] Feeding rate of the twin-screw extruder: 50 kg / h;

[0082] Characteristics of the slow-release granules:

[0083] Particle size 2 - 3 mm, length 4 - 6 mm;

[0084] Porosity of the porous structure ≥ 30% (measured by mercury intrusion method).

[0085] Step B, Composite Film Coating

[0086] Coating Liquid Preparation:

[0087] Sodium alginate (viscosity 200 - 400 mPa·s) and chitosan (degree of deacetylation ≥ 85%) are mixed in a mass ratio of 1:1;

[0088] Dissolve it in 2% acetic acid solution and stir magnetically (500 rpm) until it becomes completely transparent (about 30 minutes).

[0089] Impregnation process:

[0090] Immerse the sustained-release granules in the coating solution with a liquid-solid ratio of 5:1 (i.e., 1 kg of granules are immersed in 5 L of liquid);

[0091] The impregnation time is 5 minutes, and during this period, air is bubbled through an air pump (0.2 mPa) to prevent the deposition of granules.

[0092] Crosslinking and curing:

[0093] After impregnation, transfer the granules to 3% calcium chloride solution and let it stand for 10 minutes;

[0094] Crosslinking reaction: Ca 2+ Forms an ionic crosslinking network with sodium alginate, and chitosan enhances the membrane strength through hydrogen bonds.

[0095] Membrane thickness control:

[0096] Detect it with a microscopic thickness gauge (accuracy ±5 μm) to ensure that the membrane thickness is 50 - 80 μm.

[0097] Step C, drying treatment

[0098] Equipment: Use a hot air circulation drying oven (model DHG-9053A) to dry the crosslinked granules, with a temperature uniformity of ±1°C.

[0099] Drying parameters:

[0100] Temperature 40°C, wind speed 2 m / s;

[0101] Drying time 4 hours, and turn the granules once every 30 minutes.

[0102] Water content detection:

[0103] Use a rapid moisture analyzer (model AND MX-50), take 10 g of granules as samples, heat them to constant weight at 105°C, and calculate that the water content ≤8%.

[0104] Finished product performance verification

[0105] Sustained-release rate test:

[0106] Take 10 g of bait and place it in still water at 25°C. Take samples every 1 hour to detect the vitamin C release amount, and the cumulative release in 4 hours ≤30%.

[0107] Immune effect:

[0108] After 7 days of feeding, ELISA detects that the concentration of IgM in the serum of koi carp increases by 2.5 times (compared with uncoated bait).

[0109] By means of the combination of porous sustained-release particles and composite membrane coating technology, the release time of immune components such as vitamin C and β-glucan is extended, the intestinal absorption efficiency of koi is improved, and at the same time, water pollution caused by the rapid dissolution of bait is avoided.

[0110] 3. Citric acid buffer

[0111] Buffer premixing

[0112] Raw material ratio:

[0113] Citric acid (food grade, purity ≥ 99.5%) and sodium bicarbonate (AR grade) are accurately weighed according to a molar ratio of 1:1.5 (for example, 1 mol of citric acid + 1.5 mol of sodium bicarbonate, that is, 192 g of citric acid + 126 g of sodium bicarbonate).

[0114] Mixing process:

[0115] Use a three-dimensional motion mixer (model SYH-50) to mix citric acid and sodium bicarbonate at a rotation speed of 10 rpm for 30 minutes to ensure uniformity (CV ≤ 3%).

[0116] Sustained-release bag design and encapsulation

[0117] Bag body material: Degradable polylactic acid (PLA) film, thickness 0.1 mm, air permeability 200 mL / (m 2 ·24h·atm).

[0118] Encapsulation specifications:

[0119] Each bag is filled with 100 g of premix, with dimensions of 10 cm × 15 cm, and hot-pressed for edge sealing (sealing width ≥ 5 mm).

[0120] Each bag surface is evenly punched (hole diameter 1 mm, hole spacing 2 cm), and the total area accounts for 5%.

[0121] 4. Preparation of compound probiotic preparation

[0122] Strain ratio:

[0123] Bacillus subtilis (Bacillus subtilis, viable count ≥ 5 × 10 8 cFU / g);

[0124] Nitrobacter (Nitrosomonas spp., viable count ≥ 5 × 10 8 cFU / g);

[0125] Carrier: Wheat bran matrix (water content ≤ 10%, particle size 0.5 - 1 mm).

[0126] II. Specific steps of the treatment method:

[0127] Step 1: Early rapid intervention

[0128] Symptom monitoring and determination:

[0129] Monitoring frequency: Manually inspect the aquaculture pond three times a day, in the morning, afternoon, and evening. Focus on observing typical symptoms such as koi fish floating on the surface, lying on their sides, hyperemia on the body surface, or sunken eyeballs;

[0130] Trigger threshold: When it is found that ≥3% of the individuals show the above symptoms (for example, 30 out of 1000 are abnormal), immediately initiate the treatment procedure.

[0131] Step 2: Initiate full - process zonal management

[0132] Zonal setting:

[0133] Core area: Occupies 30% of the pond area, enclosed by a polyethylene isolation net (mesh size ≤1 cm), concentrating all diseased fish;

[0134] Buffer zone: Occupies 70% of the pond area, the activity area for healthy fish, maintaining a distance of ≥2 m from the core area.

[0135] Isolation measures:

[0136] Insert the isolation net 30 cm into the bottom silt of the pond, with a height of 50 cm above the water surface, and set PVC support piles every 2 m;

[0137] Newly diseased fish are transferred to the core area daily through a trapping net to ensure zero exposure of healthy fish.

[0138] Step 3: Preliminary treatment

[0139] Application of improved suspension:

[0140] Complete the drug application within 2 hours since the first symptom is found;

[0141] Drug preparation: Prepare an improved potassium monopersulfate compound suspension (particle size ≤20 μm) according to the suspension preparation method.

[0142] Initial dose: Evenly sprinkle at a concentration of 0.07 ppm (that is, 50 grams of drug per cubic meter of water body), covering the entire pond water surface.

[0143] Application tool: High - pressure sprayer (atomization particle size ≤100 μm), focusing on spraying the concentrated area of diseased fish.

[0144] Pre - start oxygenation equipment:

[0145] Linkage strategy: Immediately start the oxygenation equipment after applying the drug, maintaining the dissolved oxygen content ≥6 mg / L, and the initial oxygenation duration ≥4 hours per day.

[0146] Step 4: Adjust the concentration in stages

[0147] The first stage (0 - 24 hours after onset):

[0148] Drug concentration: Maintain at 0.07 ppm, apply the drug 2 times a day (with a 12 - hour interval).

[0149] Oxygenation linkage: Oxygenate for ≥2 hours each time after drug application, with a cumulative oxygenation of ≥4 hours per day.

[0150] The second stage (24 - 72 hours):

[0151] Concentration increase: Increase to 0.09 ppm, apply the drug 1 time a day (fixed at 9 am).

[0152] Oxygenation enhancement: Prolong the oxygenation duration to ≥6 hours per day, carried out in three times (2 hours each in the morning, afternoon, and night).

[0153] The third stage (after 72 hours):

[0154] Concentration decrease: Adjust to 0.05 ppm, apply the drug 1 time a day.

[0155] Feeding of immunostimulating baits: Synchronously feed immunostimulating slow - release baits (containing 0.5% vitamin C and 1.2% β - glucan), 1 time a day, for 5 consecutive days.

[0156] Probiotic input: If the ammonia nitrogen in the water body ≥0.5 mg / L, add a compound probiotic preparation (1 g / m 3 ) until the ammonia nitrogen < 0.2 mg / L;

[0157] Adding method: Premix the preparation and pond water at a ratio of 1:10, let it stand and activate for 30 minutes, then sprinkle it evenly throughout the pond;

[0158] Adding frequency: 1 time a day, continue until the ammonia nitrogen concentration < 0.2 mg / L (usually takes 3 - 5 days).

[0159] Step 5: Dynamic environmental regulation

[0160] Environmental monitoring: Record the pH value of the water body every hour. If pH > 8.0, hang a citric acid buffer slow - release bag at the water inlet of the aquaculture pond, with a slow - release rate controlled at 0.01 g / h·m 3 ; Stabilize the pH value of the water body through slow - release technology to prevent the decomposition and inactivation of potassium monopersulfate under alkaline conditions (pH > 8.0), and at the same time avoid the drastic pH fluctuation caused by the one - time addition of citric acid;

[0161] Detect the water temperature once an hour. If < 18℃, extend the dosing interval of the potassium monopersulfate compound salt suspension to once every 36 hours; Degrade the residual thiosulfate of the drug by compound probiotics, inhibit the reproduction of pathogenic bacteria (such as Aeromonas), and maintain the ecological balance of the water body.

[0162] Step 6: Treatment termination decision

[0163] Death and food intake monitoring:

[0164] Death record: No new deaths within 48 consecutive hours (the number of deaths is counted and recorded daily).

[0165] Feeding recovery: The feeding amount is restored to more than 80% of the pre-onset level (for example, the original 10kg of feed per day is restored to ≥8kg).

[0166] Viral antigen test:

[0167] Sampling method: 20 koi were randomly selected (10 in the core area and 10 in the buffer area), and body surface mucus was collected with a sterile cotton swab.

[0168] Test paper test: Use colloidal gold test paper to detect viral antigens, and the negative rate must be ≥95% (positive sample ≤1).

[0169] Gill Filament Health Assessment:

[0170] Visual observation: Check the color of the gill filaments (bright red means healthy) and their structural integrity (no ulceration or adhesion).

[0171] Microscope-assisted: Randomly take samples of gill filaments from 3 koi carps and confirm under a 40x microscope that there are no signs of parasites or bacterial infection.

[0172] Termination operation:

[0173] Conditions for drug withdrawal: no new deaths, food intake meets standards, virus-negative rate ≥ 95%, and gill filaments are healthy.

[0174] Ecological restoration: Add nitrifying bacteria for 3 consecutive days after stopping the drug to degrade drug residues and ensure that the ammonia nitrogen concentration is ≤0.2mg / L and the nitrite is ≤0.1mg / L.

[0175] 3. Data Recording and Optimization

[0176] Treatment record establishment:

[0177] Record items: including initial symptom time, application parameters (concentration, time), water quality data (pH, temperature, dissolved oxygen, ammonia nitrogen), number of deaths, food intake and virus detection results.

[0178] Database entry: Use a mobile terminal to upload to the cloud database in real time, and store it in categories such as "case table", "drug application table" and "water quality table".

[0179] Dynamic optimization strategy:

[0180] Historical data analysis: The linear regression model was used to analyze the correlation between cure rate, drug concentration, and oxygenation duration.

[0181] Parameter adjustment: If the cure rate of a certain batch is lower than 90%, the system will automatically recommend increasing the concentration by 0.02 ppm or extending the aeration time by 1 hour per day.

[0182] IV. Specific application examples

[0183] Case background: A koi fish farm (with an area of 1000 m 2 , a water depth of 1.5 m, and 5000 fish in stock) suffered an outbreak of sleeping disease, and initially 30 fish showed symptoms of floating head and body surface congestion.

[0184] Implementation process:

[0185] Early intervention:

[0186] Prepare a modified suspension (0.07 ppm) within 2 hours and sprinkle it all over the pond, and start the aeration equipment (4 hours per day);

[0187] Partition isolation: The core area (300 m 2 ) is applied with a concentration of 0.07 ppm, and the buffer area (700 m 2 ) is 0.06 ppm.

[0188] Adjustment in stages:

[0189] 24 - 72 hours: The concentration in the core area is increased to 0.09 ppm, and the aeration is 6 hours per day; the buffer area is 0.07 ppm;

[0190] After 72 hours: The concentration in the core area is reduced to 0.05 ppm, and immune bait is fed; the medicine is stopped in the buffer area;

[0191] Environmental regulation:

[0192] When pH > 8.0, hang citric acid buffer bags (0.01 g / h·m 3 ), and the pH is stabilized at 7.2;

[0193] When the ammonia nitrogen exceeds the standard (0.6 mg / L), add probiotics (100 g per day), and it drops to 0.15 mg / L after 5 days.

[0194] Treatment termination:

[0195] On the 6th day, there were no new deaths, the feed intake recovered by 85%, the negative rate of virus antigen was 96%, and the gill filaments were healthy.

[0196] Three days after stopping the medicine, ammonia nitrogen ≤ 0.1 mg / L, nitrite ≤ 0.05 mg / L.

[0197] Effect verification:

[0198] Cure rate: 95%, recurrence rate: 2%.

[0199] Drug residue: The concentration of thiosulfate is 0.007 ppm, which is lower than the fishery standard.

[0200] Cost savings: The comprehensive dosage of drugs is reduced by 18%, and the treatment cycle is shortened by 25%.

[0201] V. Summary

[0202] This embodiment provides a treatment method for sleeping disease in koi farming. Through early symptom monitoring and rapid intervention, combined with phased dynamic adjustment of drug concentration, intelligent linkage control of environmental parameters, and full-process zoning isolation management, precise drug application and ecological collaborative control are achieved; the scheme integrates improved suspension, slow-release immune bait, and probiotic technologies, taking into account the goals of efficient treatment and low residue, and improves the cure rate through a data-driven dynamic optimization strategy; the whole treatment process emphasizes environmental adaptability regulation and ecological restoration, ultimately achieving rapid virus inhibition, restoration of water ecosystem balance, and protection of healthy fish populations, and is applicable to large-scale farming scenarios.

Claims

1. A treatment method for sleeping disease in koi farming, characterized in that, It includes the following steps: Step 1: Early rapid intervention: After ≥3% of the individuals in the koi population show symptoms of floating head, lying on the side, or body surface congestion, evenly sprinkle the modified potassium monopersulfate compound salt suspension into the aquaculture water within 2 hours, with an initial concentration of 0.07 ppm, and apply it at a dose of 50 grams per cubic meter of water body; Step 2: Phased concentration adjustment: The first stage (0 - 24 hours after the onset of the disease): Maintain a concentration of 0.07 ppm and apply the medicine 2 times a day; The second stage (24 - 72 hours): Increase the concentration to 0.09 ppm, apply the medicine 1 time a day, and maintain the dissolved oxygen content ≥6 mg / L in combination with an oxygenation device; The third stage (after 72 hours): Reduce the concentration to 0.05 ppm, and simultaneously feed immune - enhancing bait containing 0.5% vitamin C once a day for 5 consecutive days; Step 3: Dynamic environmental regulation: Real-time monitor the pH value and temperature of water. When pH > 8.0, additionally add citric acid buffer (0.1 g / m 3 ) to stabilize the drug activity; When the water temperature is lower than 18°C, extend the dosing interval of the modified potassium monopersulfate compound salt suspension to once every 36 hours; Step 4: Determination of the end of treatment: When there are no new dead individuals within 48 consecutive hours and the food intake of the koi returns to more than 80% of that before the onset of the disease, stop administering the medicine.

2. The treatment method for sleeping disease in koi breeding according to claim 1, characterized in that, The preparation method of the modified potassium monopersulfate compound salt suspension is as follows: Mix AR - grade potassium monopersulfate compound salt and food - grade diatomaceous earth in a mass ratio of 1:0.2, add 0.05% polyethylene glycol (PEG - 4000) as a dispersant, and process it through a high - speed shear emulsifier for 10 minutes to form a uniform suspension with a particle size ≤20 μm.

3. A treatment method for sleep disease in koi farming according to claim 1, characterized in that, The usage strategy of the oxygenation device is as follows: During the medicine application period in the second stage (24 - 72 hours), start the oxygenation device after each medicine application every day to maintain the dissolved oxygen content ≥6 mg / L, and the oxygenation duration is positively correlated with the drug concentration. Specifically: When the concentration is 0.07 ppm, the oxygenation duration ≥4 hours per day; When the concentration is 0.09 ppm, the oxygenation duration ≥6 hours per day.

4. A treatment method for sleep disease in koi farming according to claim 1, characterized in that, The preparation method of the immune - enhancing bait is as follows: Step A, preparation of the base material: Using fish meal as the basic raw material, add 0.5% vitamin C, 1.2% β - glucan, and 0.3% allicin, and use a twin - screw extruder to make slow - release granules with a porous structure at 120 - 130°C and a pressure of 30 - 40 MPa. The particle size of the slow - release granules is 2 - 3 mm; Step B, composite film coating: Immerse the slow - release granules in a sodium alginate - chitosan composite solution (the mass ratio of sodium alginate to chitosan is 1:1, and the solvent is 2% acetic acid solution), take them out after impregnation for 5 minutes, and place them in 3% calcium chloride solution for cross - linking and curing for 10 minutes to form a composite film with a thickness of 50 - 80 μm; Step C, drying treatment: Hot - air dry the coated granules at 40°C until the water content ≤8%.

5. A treatment method for sleeping disease in koi farming according to claim 1, characterized in that, The addition method of the citric acid buffer is as follows: Citric acid and sodium bicarbonate are premixed at a molar ratio of 1:1.5 and then encapsulated in a degradable slow-release bag, which is suspended at the inlet of the aquaculture pond, and the slow-release rate is controlled at 0.01 g / h·m 3 .

6. A treatment method for sleep disease in koi breeding according to claim 1, characterized in that, In step 3, when the ammonia nitrogen concentration in the water body ≥ 0.5 mg / L, a compound probiotic preparation (containing Bacillus subtilis and Nitrobacter, viable count ≥ 1×10 8 CFU / g) is synchronously added, with a dosage of 1 g / m 3 , once a day, continuing until the ammonia nitrogen concentration < 0.2 mg / L.

7. A treatment method for sleeping disease in koi farming according to claim 1, characterized in that, The determination of the end of treatment also includes: Collect mucus samples from the body surface of the koi, use the test paper method to detect that the negative rate of virus antigen ≥95%, and there is no visible congestion or ulcer in the gill filament tissue.

8. A treatment method for sleep disease in koi breeding according to claim 1, characterized in that, Adopt zoned isolation management throughout the treatment process: Divide the disease occurrence pond into a core area (where diseased fish concentrate) and a buffer area (where healthy fish move). Increase the drug application concentration in the core area by 10%, maintain the basic concentration in the buffer area, and set up a physical isolation net (mesh size ≤ 1 cm) between the two areas.

9. A treatment method for sleep disease in koi breeding according to claim 1, characterized in that, Establish a treatment record database: Record the initial symptom time, drug application parameters, water quality changes, and cure rate data for each batch of treatment. Optimize the concentration gradient through statistical analysis to increase the cure rate from 90% to 93% - 95%.

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