Method and system for diagnosing and improving sub-health of aquatic animals based on secondary strike theory

Abstract

The invention belongs to the technical field of aquaculture, and discloses a method for diagnosing and improving sub-health of aquatic animals based on a secondary strike theory, which systematically summarizes threats of feed to health into two levels of'nutrition 'and'non-nutrition' for the first time, and clarifies a synergistic / progressive action mechanism of the feed. Predictability is achieved, early diagnosis and early warning of the health risk of the aquatic animals are achieved, and passive disease treatment is changed into active health care. Due to guidance, a complete and operable solution from diagnosis to intervention is provided, and production practice is directly guided. And differentiated intervention strategies are provided according to different risk levels, so that the cost is effectively reduced, and the prevention and control efficiency is improved.

Description

Technical Field

[0001] This invention belongs to the field of aquaculture technology, and in particular relates to a method and system for diagnosing and improving the sub-health of aquatic animals based on the theory of secondary impact. Background Technology

[0002] In the current aquaculture industry, farmers mainly rely on experience and observation to judge the health status of aquatic animals, and usually only take treatment measures when disease symptoms are obvious. This passive "treat the disease as it is" model stems from limited understanding of the process of health deterioration, and its deficiency lies in the inability to provide early warning and intervention for potential, gradual health risks.

[0003] Ultimately, aquaculture can be viewed as a process of transforming feed ingredients into aquatic products using aquatic animals as a biological factory. Feed, as the core raw material, directly determines the long-term health and operational efficiency of this "factory." This raw material can be systematically divided into two categories: nutrients and non-nutrients. Its impact on the organism is not isolated but rather a continuous, time-sequential, and synergistic process.

[0004] However, existing cognitive and technological systems have failed to reveal this underlying mechanism:

[0005] First, there is the insidious "first blow." An imbalance of nutrients (such as fat and protein) in feed, or overfeeding, is like supplying the "factory" with the wrong proportions of fuel and building materials, causing its metabolic system to operate under overload, leading to sub-health conditions such as excessive fat deposition and impaired liver function. This problem is insidious and difficult to quantify and detect in daily observation.

[0006] Then comes the fatal "second blow." When the "factory" is weakened by the first blow, the continuous intrusion of non-nutritive substances in the feed (such as mycotoxins and oxidation products) will cause cumulative damage to the already damaged liver, pancreas, and other critical "equipment." This second blow often becomes the last straw that breaks the camel's back. When the external environment deteriorates, such as water quality deteriorates or pathogens increase, it causes the "factory" to shut down—that is, a disease outbreak.

[0007] Therefore, the fundamental contradiction currently facing the industry is that although practitioners have the desire to maintain their health, existing technologies lack a theoretical model that can reveal the inherent law of "the phased and synergistic effects of nutritional and non-nutritional stress," which makes it impossible to achieve systematic diagnosis, early warning, and precise intervention based on risk levels for sub-health status from the source. Summary of the Invention

[0008] To address the problems existing in the prior art, this invention provides a method for diagnosing and improving the sub-health of aquatic animals based on the theory of secondary impact.

[0009] This invention is implemented as follows: A method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory includes:

[0010] Step 1, assess the risk of a "first strike";

[0011] Step 2, assess the risk of a "second strike";

[0012] Step 3: Comprehensive health risk assessment;

[0013] Step 4: Tiered intervention strategy.

[0014] Furthermore, the assessment of the "first strike" risk:

[0015] Analyze feed formulations, test the composition and proportion of its nutrients, and determine whether there is a risk of sub-health problems such as fatty liver and obesity.

[0016] Furthermore, the assessment of the "second strike" risk:

[0017] The content of key non-nutritive substances in feed is detected to assess their potential toxicity to organs such as the liver and pancreas of fish.

[0018] Furthermore, the comprehensive health risk rating:

[0019] Based on the test results from steps one and two, and combined with the clinical signs of the fish, a health risk rating system was established.

[0020] Furthermore, the tiered intervention strategy:

[0021] Implement corresponding intervention measures based on different health risk ratings:

[0022] High-risk groups: Immediately stop using the existing feed, replace it with a precisely formulated health feed, and adjust the water quality.

[0023] For medium-risk groups: adjust feeding strategies and add specific feed additives to existing feed to mitigate the impact;

[0024] Low-risk groups.

[0025] Furthermore, for the aforementioned low-risk groups: suggestions for feeding strategies or feed formulation optimization are proposed to prevent the occurrence of "attacks" from the source.

[0026] Another objective of this invention is to provide a system for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory, comprising:

[0027] The risk assessment module is used to assess the risk of a "first strike" and the risk of a "second strike".

[0028] The rating module is used for comprehensive health risk rating.

[0029] The intervention module is used for tiered intervention strategies.

[0030] Another object of the present invention is to provide a computer device, the computer device including a memory and a processor, the memory storing a computer program, the computer program being executed by the processor causing the processor to perform the steps of the method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory.

[0031] Another object of the present invention is to provide a computer-readable storage medium storing a computer program, which, when executed by a processor, causes the processor to perform the steps of the method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory.

[0032] Another objective of this invention is to provide an information data processing terminal for implementing the system for diagnosing and improving the sub-health of aquatic animals based on the secondary impact theory.

[0033] Based on the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solution to be protected by this invention are as follows:

[0034] This study systematically categorizes the health threats posed by feed into two levels: "nutritional" and "non-nutritional," and elucidates their synergistic / progressive mechanisms.

[0035] Its predictive capabilities enable early diagnosis and warning of health risks in aquatic animals, transforming "passive disease treatment" into "proactive health care."

[0036] It provides guidance and offers a complete and actionable solution from diagnosis to intervention, directly guiding production practices.

[0037] Precision allows for differentiated intervention strategies based on different risk levels, effectively reducing costs and improving prevention and control efficiency.

[0038] (1) The expected benefits and commercial value of the technical solution of the present invention after transformation are as follows: The technical solution of the present invention has significant commercial value after transformation. Its core lies in transforming the innovative "secondary strike" theory into a feasible health management solution, providing early warning and precise intervention services for farmers, which can effectively reduce breeding risks and reduce disease losses. At the same time, it provides technical support for feed companies to develop high-value-added functional products and supporting services, creating new profit growth points.

[0039] (2) The technical solution of the present invention fills the technical gap in the industry at home and abroad: The present invention fills the technical gap in the industry at home and abroad, and for the first time establishes a systematic "secondary attack" theoretical model and supporting operation method, which solves the long-standing technical problem of lack of a health management system in the aquaculture field that can simultaneously integrate nutritional and non-nutritive factors and elucidate their synergistic harm mechanism.

[0040] (3) Whether the technical solution of the present invention solves the technical problem that people have been eager to solve but have never been able to solve: The present invention successfully solves the technical problem that people have been eager to solve but have never been able to break through, namely, how to achieve early identification and effective intervention of health risks before obvious clinical symptoms appear in aquatic animals, realizing a fundamental shift from passive "treatment after disease" to proactive "prevention before disease".

[0041] (4) Whether the technical solution of the present invention overcomes technical bias: The present invention overcomes technical bias in the industry, breaks the traditional limited thinking of viewing nutrition and toxin problems in isolation, innovatively reveals the intrinsic connection and progressive harm mechanism between the two, and establishes a graded intervention strategy based on risk level, changing the previous single and extensive prevention and control model. Attached Figure Description

[0042] Figure 1 This is a flowchart of a method for diagnosing and improving the sub-health of aquatic animals based on the secondary impact theory, provided in an embodiment of the present invention.

[0043] Figure 2 This is a flowchart of the method for assessing the risk of "first strike" provided in an embodiment of the present invention.

[0044] Figure 3 This is a flowchart of a method for assessing the risk of a "second strike" provided in an embodiment of the present invention.

[0045] Figure 4 This is a system structure diagram of aquatic animals for diagnosing and improving sub-health based on the secondary impact theory, provided in an embodiment of the present invention.

[0046] Figure 5 The following are typical liver, duodenum, and valvular intestine tissue sections of hybrid sturgeon provided in this embodiment of the invention: Figures A1, A4, and A7 show liver, duodenum, and valvular intestine sections of the control group; Figures A2, A5, and A8 show liver, duodenum, and valvular intestine sections of the HCL group; Figures A3, A6, and A9 show liver, duodenum, and valvular intestine sections of the HCLH group. In the liver sections, hexagons indicate hepatocyte fusion, and triangles indicate inflammatory cell infiltration; in the intestinal sections, ellipses indicate pancreatic islets, stars indicate intestinal mucosal fusion, arrows indicate intestinal mucosal hyperplasia, circles indicate intestinal mucosal necrosis, and triangles indicate vacuolar images of the serosa or muscle layer.

[0047] Figure 6The images provided in this embodiment of the invention show typical liver, duodenum, and valvular tissue sections of hybrid sturgeon after intervention. Figures B1, B4, and B7 show liver, duodenum, and valvular sections from the control group; Figures B2, B5, and B8 show liver, duodenum, and valvular sections from the CJ group; and Figures B3, B6, and B9 show liver, duodenum, and valvular sections from the CB group. In the liver sections, triangles indicate inflammatory cell infiltration.

[0048] Figure 7 This is an image of a grass carp provided in an embodiment of the present invention.

[0049] Figure 8 This is a grass carp liver tissue section provided in an embodiment of the present invention. Arrows indicate sinusoidal congestion, circles indicate hepatocyte fusion, and triangles indicate inflammatory cell infiltration.

[0050] Figure 9 This is an image of a grass carp provided in an embodiment of the present invention.

[0051] Figure 10 This is a slice of grass carp liver tissue provided in an embodiment of the present invention. Arrows indicate sinusoidal congestion, circles indicate hepatocyte fusion, and triangles indicate liver tissue damage.

[0052] Figure 11 The arrow in the anatomical diagram of the Chinese soft-shelled turtle provided in this embodiment of the invention indicates the fat layer.

[0053] Figure 12 The oval image shows HE slices of the intestine and liver of the Chinese soft-shelled turtle provided in this embodiment of the invention, indicating damage to the intestinal villi, and the arrow indicates venous sinus congestion.

[0054] Figure 13 The arrow in the anatomical diagram of the Chinese soft-shelled turtle provided in this embodiment of the invention indicates the fat layer.

[0055] Figure 14 This is an oval image showing intestinal villus damage in HE slices of the intestine and liver of the Chinese soft-shelled turtle provided in an embodiment of the present invention. Detailed Implementation

[0056] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.

[0057] In current aquaculture practices, sub-health conditions are usually not caused by a single factor, but rather are the result of a gradual accumulation of the combined effects of nutritional imbalance and environmental stress. Traditional diagnostic methods mainly rely on clinical symptom observation and single-factor biochemical index analysis, neglecting the interaction between feed nutrient composition and potential toxicological factors, making it difficult to detect hidden risks in a timely manner. This invention introduces the two-hit theory, viewing the occurrence of sub-health in aquatic animals as a process jointly triggered by a first metabolic load shock and a second toxicological stress shock. It reconstructs the risk identification mechanism from a systems physiology perspective, enabling early warning and precise intervention.

[0058] In the initial risk assessment phase, the method analyzes the metabolic burden of energy and nutrient supply by examining the proportions of protein, lipids, carbohydrates, and trace elements in the feed. This phase focuses on identifying fat deposition, oxidative stress, and hepatopancreatic metabolic disorders caused by excessive lipids or unbalanced amino acids in the feed. By establishing a nutritional metabolic risk model, feed formulation characteristics are mapped to hepatic lipid metabolism pathways, antioxidant enzyme systems, and inflammatory signaling networks, thereby achieving a quantitative assessment of metabolic impact.

[0059] The second risk assessment focuses on monitoring potential non-nutritive factors in feed and water, including mycotoxins, oxidation products, environmental heavy metal residues, and byproducts of artificial additives. This process uses multi-omics analysis combined with toxicological parameters to estimate the risk of damage to key organs. With the hepatocellular carcinoma and pancreas as the core target organs, a toxicological stress index is established using biochemical markers such as ALT, AST, MDA, and antioxidant enzyme activity indicators to achieve early identification of latent subclinical toxicities.

[0060] The comprehensive health risk rating process, based on the quantitative results of the first and second exposures, combines surface symptoms, feeding behavior, and metabolic product characteristics to construct a multidimensional health assessment matrix. This matrix reflects the degree of damage to each physiological system through a weighted model, outputting a graded health risk index to provide a basis for intervention strategy decisions. This process embodies the combination of physiological information fusion and risk prediction algorithms, enabling dynamic adjustment of assessment thresholds to adapt to different species and farming environments.

[0061] The intervention strategy module develops tiered response plans based on health risk assessment results. Management of high-risk groups focuses on source control and physiological repair, restoring metabolic homeostasis through switching to low-fat, antioxidant-fortified feed and adjusting water physicochemical parameters. Medium-risk groups utilize a combination of optimized feeding and functional additives to achieve metabolic buffering, with a focus on hepatoprotective and detoxifying substances to mitigate cumulative stress. Low-risk groups prioritize prevention at the source, using nutritional optimization and feed ingredient substitution to inhibit the formation of potential stressors.

[0062] The entire system operates on a closed-loop feedback mechanism, with risk assessment as input, health rating as the core, and intervention and control as output. The risk assessment and rating modules share data to achieve iterative model updates, while the intervention module dynamically optimizes the intensity and timing of interventions based on historical feedback, enabling full-process control from diagnosis to improvement. This method breaks through the traditional experience-based judgment model, providing a quantifiable, traceable, and controllable scientific basis for the identification and intervention of sub-health in aquatic animals, and possesses significant potential for industrial application.

[0063] like Figure 1 As shown in the figure, an embodiment of the present invention provides a method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory, which includes the following steps:

[0064] S101, assess the risk of a "first strike";

[0065] S102, assess the risk of a "second strike";

[0066] S103, Comprehensive Health Risk Rating;

[0067] S104, Tiered Intervention Strategy.

[0068] like Figure 2 As shown, the embodiments of the present invention provide an assessment of the "first strike" risk:

[0069] S201 involves analyzing feed formulations, testing the composition and proportion of nutrients, and determining whether there is a risk of sub-health problems such as fatty liver and obesity.

[0070] like Figure 3 As shown, the embodiments of the present invention provide an assessment of the risk of a "second strike":

[0071] S301 is used to detect the content of key non-nutritive substances in feed and assess their potential toxicity to organs such as the liver and pancreas of fish.

[0072] The comprehensive health risk rating provided by the embodiments of the present invention:

[0073] Based on the test results from steps one and two, and combined with the clinical signs of the fish, a health risk rating system was established.

[0074] The tiered intervention strategy provided in this embodiment of the invention:

[0075] Implement corresponding intervention measures based on different health risk ratings:

[0076] High-risk groups: Immediately stop using the existing feed, replace it with a precisely formulated health feed, and adjust the water quality.

[0077] For medium-risk groups: adjust feeding strategies and add specific feed additives to existing feed to mitigate the impact;

[0078] Low-risk groups.

[0079] The low-risk groups provided by this invention include suggestions for feeding strategies or feed formulation optimization to prevent the occurrence of "attacks" from the source.

[0080] like Figure 4 As shown in the figure, an embodiment of the present invention provides a system for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory, comprising:

[0081] The risk assessment module is used to assess the risk of a "first strike" and the risk of a "second strike".

[0082] The rating module is used for comprehensive health risk rating.

[0083] The intervention module is used for tiered intervention strategies.

[0084] Another object of the present invention is to provide a computer device, the computer device including a memory and a processor, the memory storing a computer program, the computer program being executed by the processor causing the processor to perform the steps of the method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory.

[0085] Another object of the present invention is to provide a computer-readable storage medium storing a computer program, which, when executed by a processor, causes the processor to perform the steps of the method for diagnosing and improving the sub-health of aquatic animals based on the two-hit theory.

[0086] Another objective of this invention is to provide an information data processing terminal for implementing the system for diagnosing and improving the sub-health of aquatic animals based on the secondary impact theory.

[0087] Specific implementation of the present invention:

[0088] This invention aims to overcome the shortcomings of existing technologies and provide a method for early warning, systematic diagnosis, and precise intervention of aquatic animal health problems caused by feed. Based on an innovative "secondary impact" theoretical model, this method can fundamentally reduce aquaculture risks and improve aquaculture efficiency.

[0089] Technical solution:

[0090] The core of the method described in this invention lies in the application of the "secondary strike" theoretical model, which systematically divides the threat of feed to fish into two stages.

[0091] The first setback is caused by an imbalance or excess of nutrients in the feed. For example, an excessively high fat / protein ratio or excessive carbohydrates can lead to metabolic disorders in the fish (such as fat deposition in liver cells and decreased function), resulting in a "sub-healthy" state.

[0092] The second blow is caused by the continuous intake of non-nutritive substances in the feed. Examples include mycotoxins, oil oxidation products (malondialdehyde), and histamine. When the fish's detoxification organs, such as the hepatopancreas, are damaged due to the "first blow," their ability to detoxify these non-nutritive substances decreases, leading to toxin accumulation, organic damage, and ultimately, disease outbreaks.

[0093] The technical solution of the present invention includes the following steps:

[0094] A. Diagnosis and Assessment Phase

[0095] Step 1 (Assessing the risk of a "first strike"):

[0096] Analyze feed formulations to test the composition and proportion of nutrients (such as fat, protein, and carbohydrates) to determine whether there is a risk of sub-health problems such as fatty liver and obesity.

[0097] Step Two (Assessing the Risk of a "Second Strike"):

[0098] The content of key non-nutritive substances (such as aflatoxin B1, malondialdehyde, histamine, etc.) in feed is detected to assess their potential toxicity to organs such as the liver and pancreas of fish.

[0099] Step 3 (Comprehensive Health Risk Assessment):

[0100] Based on the test results of Step 1 and Step 2, and combined with the clinical signs of the fish (such as liver color and body shape), a health risk rating system (e.g., low risk, medium risk, high risk) is established.

[0101] B. Interventions and Solutions

[0102] Step Four (Tiered Intervention Strategy):

[0103] Implement corresponding intervention measures based on different health risk ratings:

[0104] High-risk groups: Immediately stop using the existing feed, adjust the feeding strategy, and replace it with a precisely formulated health feed (such as a low-fat, high-protein feed with sufficient bile acids and liver-protecting and detoxifying Chinese medicine), and carry out water quality control.

[0105] For medium-risk groups: Adjust feeding strategies and add specific feed additives (such as probiotics, liver-protecting and detoxifying traditional Chinese medicine, and antioxidants) to the existing feed to mitigate the "hit".

[0106] For low-risk groups: propose feeding strategies or feed formulation optimization suggestions to prevent the "attack" from occurring at the source.

[0107] I. Specific application areas or related products of this invention.

[0108] Example 1: Applying the described method to diagnose sub-health status in sturgeon and verify the "secondary blow" model

[0109] This embodiment uses juvenile hybrid sturgeon (Acipenser schrenckii ♀ × A. baeri ♂) as the subject to demonstrate how to apply the method described in this invention to diagnose their sub-health state caused by feed and to verify the scientific validity of the "secondary impact" theoretical model.

[0110] 1. Diagnosis and Assessment Phase

[0111] The induction and confirmation of the first strike (nutritional strike) and the manifestation of the second strike (non-nutritional strike).

[0112] 1.1 Operation:

[0113] The experimental fish were divided into three groups. One group (Group C) was fed a nutritionally balanced control diet, while the other group (HCL group) was fed a high-sugar, high-fat diet (fat level 17.83%, sugar level 22%, denoted as HCL) and a high-sugar, high-fat diet (fat level 17.83%, sugar level 22%, histamine 2000mg / kg, denoted as HCLH) made from purified raw materials such as casein and gelatin. The fish were cultured for 6 weeks.

[0114] 1.2 Diagnostic Results:

[0115] Body shape indicators: As shown in Table 1, the visceral-to-body ratio, liver-to-body ratio and fatness of the HCL and HCLH groups were significantly increased (P < 0.05), indicating increased visceral fat deposition.

[0116] Table 1 Body Shape Indicators

[0117] Note: Data in the same column with different lowercase subscripts indicate significant differences between groups (P < 0.05).

[0118] Body composition: As shown in Table 2, compared with group C, the fat content in different parts of HCL and HCLH groups was significantly increased (P < 0.05), with the liver fat content reaching more than 40% of wet weight. This confirms that the liver, as a core fat storage organ, has experienced severe nutritional fat deposition. Moreover, the fat content in the abdominal muscles, liver, and internal organs of the HCLH group was significantly higher than that of other groups (P < 0.05), indicating that the presence of non-nutritive substances further promotes fat deposition.

[0119] Table 2 Fat deposition in different locations

[0120]

[0121] Note: Data in the same column with different lowercase subscripts indicate significant differences between groups (P < 0.05).

[0122] Histology: The sturgeon's liver is also a fat storage organ, therefore, by Figure 5 As can be seen, all groups showed hepatocyte vacuolation in HE sections, but the HCL group showed cell fusion and inflammatory cell infiltration, which was more severe in the HCLH group. Intestinal sections also showed that the HCL and HCLH groups had more severe intestinal mucosal fusion, intestinal mucosal hyperplasia, intestinal mucosal necrosis, and vacuolation of the serosa or muscle layer.

[0123] 1.3 Conclusion:

[0124] Hybrid sturgeon fed HCL feed are at a "medium risk" level, while those fed HCLH are at a "high risk" level. The root cause of their health problems lies in the severe fatty liver and metabolic disorders caused by the imbalance of high sugar and high fat nutrition (the first blow), which has significantly weakened the fish's health foundation. When faced with the "second blow" of the non-nutritive substance histamine, more obvious tissue damage occurs.

[0125] 2. Intervention Solutions (Theoretical Verification)

[0126] 2.1 Intervention Strategies:

[0127] In this embodiment, three groups were set up. The hybrid sturgeon that were originally fed Group C feed continued to be fed Group C feed and were still named Group C. The experimental fish that were originally fed HCl feed adopted the feeding strategy of "intermittent feeding of Group C feed" and were named Group CJ. This theoretically verified that controlling the "first blow" can effectively block the chain of health deterioration. The hybrid sturgeon that were originally fed HClH feed were fed a reduced amount of Group C feed and mixed with bile acids and liver-protecting and detoxifying Chinese medicine, and were named Group CB. This theoretically verified that controlling the "second blow" can effectively block the chain of health deterioration.

[0128] 2.2 Intervention cycle:

[0129] It lasted for 6 weeks.

[0130] 2.3 Validation of Intervention Effect

[0131] Body shape indicators: As shown in Table 3, after six weeks, there were no significant differences in the ratio of organs to body mass, the ratio of liver to body mass, and the degree of fatness among the groups (P > 0.05), indicating that the intervention measures achieved an effect on appearance.

[0132] Table 3 Body Shape Indicators

[0133]

[0134] Body composition: As shown in Table 4, after six weeks, there was no significant difference between CJ and the control group (P > 0.05), while the fat content in the head and liver of the CB group was still higher than that of the control group (P < 0.05), indicating that the effects of the second blow from non-nutritive substances still existed.

[0135] Table 4 Fat content in different body parts

[0136]

[0137] Note: Data from the same group with different lowercase subscripts indicate significant differences between groups (P < 0.05).

[0138] Histology: by Figure 6 It is evident that the liver and intestines of both the CJ and CB groups improved after the intervention, but inflammatory cell infiltration was still found in the liver of the CB group, indicating that the intervention can improve the health of the hybrid sturgeon.

[0139] 3. Conclusion

[0140] This embodiment fully demonstrates the occurrence of the "secondary blow" in hybrid sturgeon and the outcome after intervention. By applying the method of this invention, the sub-health state of hybrid sturgeon caused by "nutritional excess" (first blow) and non-nutritional substances (second blow) can be accurately diagnosed. Controlled experiments demonstrate that by managing feeding strategies and nutritional strategies, the first blow can be controlled, and the second blow can be effectively prevented and alleviated, thereby maintaining the health of the fish. This proves the scientific validity, predictive power, and significant application value of the method of this invention in aquaculture.

[0141] Example 2: Applying the described method to diagnose and improve the sub-health state of grass carp

[0142] This embodiment demonstrates the entire process of diagnosing and implementing precise intervention for the sub-health status of grass carp in a fish farm using the method described in this invention.

[0143] 1. Diagnosis and Assessment Phase

[0144] (1) Sample collection and initial risk assessment

[0145] At a grass carp farm, ponds with similar farming conditions were selected, and samples of a commercial feed (already fed for 30 days) and representative grass carp were collected. Through on-site observation, some fish were found to exhibit potential sub-health signs such as abdominal distension and pale liver color. Figure 7 ).

[0146] (2) Risk assessment of the first strike

[0147] Nutritional analysis of the feed is shown in Table 5. The results show that although the feed can meet the basic nutritional requirements, there is a potential risk of imbalance in carbohydrate, fat and protein ratios, which may lead to excessive energy metabolic load in grass carp and constitute a potential "first blow".

[0148] Table 5. Nutritional and non-nutritive content of a certain commodity feed

[0149]

[0150] (3) Risk assessment of the second strike

[0151] Non-nutritional substance testing of the same batch of feed did not reveal any serious exceedances of specific toxins, but aflatoxin B1, malondialdehyde, and histamine, which are of key concern, were all detected (Table 5). In particular, the content of aflatoxin B1 was close to the upper limit of GB13078-2017 "Feed Hygiene Standard". Furthermore, subsequent biochemical and histological analysis of fish serum revealed signs of oxidative stress, indicating that the fish were in an internal metabolic stress state, thus indicating a "second blow".

[0152] (4) Comprehensive health risk rating

[0153] Precise laboratory tests were performed on the fish, and the results are as follows:

[0154] Liver health: Grass carp not only have whitish livers, but also high serum triglyceride (TG) levels (5.27 mmol / L). Figure 8 As can be seen, the liver tissue section showed sinusoidal congestion, partial hepatocyte fusion, and inflammatory cell infiltration, which are typical manifestations of fatty liver and liver damage.

[0155] (5) Comprehensive judgment

[0156] The grass carp in this pond are at a "medium risk" level. The root cause of their health problems lies in the metabolic disorders and fatty liver caused by nutritional imbalance (the first blow), which has significantly weakened the fish's health foundation. This has led to obvious tissue damage and functional decline in their liver and intestines when faced with non-nutritive substances (aflatoxin B1, malondialdehyde, histamine, etc., the second blow).

[0157] 2. Intervention and Solutions

[0158] Based on the determination of "medium risk" level, immediately implement the corresponding tiered intervention strategy:

[0159] Intervention measures:

[0160] (1) Immediately add functional feed additives such as bile acids, liver-protecting and detoxifying Chinese medicine, and lysophospholipids to the feed. These are compound substances that promote fat metabolism, protect liver and gallbladder health, and enhance antioxidant capacity. At the same time, use potassium persulfate to adjust the water and improve the bottom, and spray Bacillus spores at noon on sunny days, once every 5 days.

[0161] (2) Intervention cycle

[0162] Feed continuously for 28 days.

[0163] (3) Validation of intervention effect

[0164] After the intervention, indicators were tested on the grass carp, and the results confirmed that the intervention was effective.

[0165] Improvement of obesity and fat deposition: Compared with the control group without intervention, the fatness, liver-to-body ratio and viscera-to-body ratio of the intervention group were significantly reduced (Table 6), and the body shape of the fish returned to normal (P<0.05).

[0166] Table 6 Body Shape Indicators

[0167]

[0168] Note: Different letters in the same column indicate significant differences (P < 0.05).

[0169] Restore liver health:

[0170] Serum triglyceride (TG) levels decreased significantly to 1.89 mmol / L, indicating a significant improvement in lipid metabolism.

[0171] Depend on Figure 9 As can be seen, the liver color of the intervention group turned into a healthy dark red compared to the non-intervention group.

[0172] Depend on Figure 10 As can be seen, liver tissue sections showed that the hepatocyte structure was compact and intact, and the congestion was basically gone. However, in the uninterventional group, the congestion of the hepatic sinusoids was aggravated, some hepatocytes fused, and liver tissue damage occurred.

[0173] 3. Conclusion

[0174] This embodiment fully demonstrates the entire process from "risk diagnosis" to "precise intervention" and then to "effect verification." By applying the method of this invention, the sub-health state of grass carp caused by the first "nutrient" attack and the second "non-nutrient" attack was accurately diagnosed. By feeding targeted functional additives and improving water quality, the sub-health state was successfully reversed, and liver and intestinal health was significantly improved, proving the effectiveness and great value of the method of this invention in production practice.

[0175] Example 3: Application of the described method to diagnose and improve the sub-health state of Chinese soft-shelled turtles.

[0176] 1. Diagnosis and Assessment Phase

[0177] (1) Sample collection and initial risk assessment

[0178] At a Chinese soft-shelled turtle farm, ponds with similar farming conditions were selected, and samples were collected from the commercial feed being used (already fed for 60 days) and representative samples of Chinese soft-shelled turtles. Through on-site observation, the turtles showed normal body color and normal, healthy red liver color, but slightly higher internal fat content. Figure 11 ).

[0179] Table 7. Nutritional and Non-Nutritional Content of a Certain Commodity Feed

[0180]

[0181] (2) First strike risk assessment

[0182] Nutritional analysis of the feed is shown in Table 7. The results show that the feed can meet basic nutritional needs, and the ratio of carbohydrates, fats and proteins is relatively balanced. Fat accumulation is likely related to the large feeding amount, which may constitute a potential "first hit".

[0183] (3) Risk assessment of the second strike

[0184] Non-nutritional substance tests were conducted on the same batch of feed, as shown in Table 7. The tests on volatile basic nitrogen, malondialdehyde, and histamine in the feed that may affect the health of Chinese soft-shelled turtles revealed that the contents were all relatively low, indicating a low possibility of a "secondary impact".

[0185] (4) Comprehensive health risk rating

[0186] Precise laboratory tests and histological observations were performed on the turtle body, and the results were as follows: Figure 12 :

[0187] Liver and intestinal health: The structure of the intestines and liver is clear and intact, but there is slight damage to the intestinal villi, and the liver cells have vacuoles with slight sinus congestion.

[0188] (5) Comprehensive judgment

[0189] This group of Chinese soft-shelled turtles is at a "low-risk" level. The root cause of their health problems lies in the fact that the feeding amount was slightly too large (the first shock), which led to an increased metabolic load and insufficient antioxidant capacity, resulting in functional and structural damage to the liver when coping with daily metabolic stress.

[0190] 2. Intervention and Solutions

[0191] Based on the determination of "low risk" level, immediately implement the corresponding tiered intervention strategy:

[0192] (1) Intervention measures

[0193] Reduce the amount of food given according to the original feeding plan by 3%-5% to reduce fat accumulation.

[0194] (2) Intervention cycle

[0195] Feed continuously for 60 days.

[0196] (3) Validation of intervention effect

[0197] After the intervention, tests were conducted, such as... Figure 13 The results confirmed that the intervention was effective.

[0198] On-site observation revealed a reduction in internal fat, and inquiries confirmed that the feed conversion ratio remained at a certain level.

[0199] A value of around 1.1 indicates that growth was not affected.

[0200] Precise laboratory tests and histological observations were performed on the turtle body, and the results were as follows: Figure 14 :

[0201] Liver and intestinal health: The structure of the intestines and liver is clear and intact, but compared with before the intervention, the damage to the intestinal villi is less severe, and there are vacuoles in the liver cells, but the venous sinus congestion has disappeared, indicating that the Chinese soft-shelled turtle is in good health.

[0202] 4. Conclusion

[0203] This embodiment once again fully demonstrates the entire process from "risk diagnosis" to "precision nutritional intervention" and then to "effect verification." By applying the method of this invention, the health of the Chinese soft-shelled turtle was accurately diagnosed as being at low risk, and through intervention, the effectiveness and application value of the method of this invention in the practice of healthy Chinese soft-shelled turtle farming were proven.

[0204] II. Evidence related to the technical effects obtained by the embodiments of the present invention.

[0205] It should be noted that embodiments of the present invention can be implemented in hardware, software, or a combination of both. The hardware portion can be implemented using dedicated logic; the software portion can be stored in memory and executed by a suitable instruction execution system, such as a microprocessor or dedicated-design hardware. Those skilled in the art will understand that the above-described devices and methods can be implemented using computer-executable instructions and / or included in processor control code, for example, such code provided on a carrier medium such as a disk, CD, or DVD-ROM, a programmable memory such as read-only memory (firmware), or a data carrier such as an optical or electronic signal carrier. The devices and modules of the present invention can be implemented by hardware circuitry such as very large-scale integrated circuits or gate arrays, semiconductors such as logic chips, transistors, or programmable hardware devices such as field-programmable gate arrays, programmable logic devices, etc., or by software executed by various types of processors, or by a combination of the above-described hardware circuitry and software, such as firmware.

[0206] The above description is merely a specific embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any modifications, equivalent substitutions, and improvements made by those skilled in the art within the scope of the technology disclosed in the present invention, and within the spirit and principles of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for diagnosing and improving sub-health conditions in aquatic animals based on the two-hit theory, characterized in that, Includes the following steps: (1) Conduct the first risk assessment, test the proportion of nutrients in the feed formula, and calculate the metabolic load of lipids, proteins and carbohydrates; (2) Conduct a second risk assessment, detect the content of non-nutritive compounds in the feed and assess their toxicological risk to the hepatopancreatic tissue; (3) Based on the combined results of the first and second risk assessments, establish a health risk rating model and output a health risk index; (4) Implement graded intervention measures based on the health risk index to diagnose and improve the sub-health status of aquatic animals.

2. The method as described in claim 1, characterized in that, The first strike risk assessment was conducted by establishing a nutritional metabolic load model and using energy metabolism rate, lipid deposition coefficient and oxidative stress index to calculate metabolic risk value.

3. The method as described in claim 1, characterized in that, The second strike risk assessment calculates the toxicological risk value based on the product of organ toxicity coefficient and exposure dose by detecting the concentrations of mycotoxins, heavy metal residues, and oxidation byproducts.

4. The method as described in claim 1, characterized in that, The health risk rating model generates a health risk index based on the weighted sum of the risk values ​​of the first and second strikes, with the weighting coefficients dynamically adjusted according to the fish species, age, and ambient temperature.

5. The method as described in claim 1, characterized in that, The tiered intervention measures include: High-risk groups should undergo feed replacement and water body physicochemical regulation. For medium-risk groups, adjust feed feeding strategies and intervene with functional additives; Feed formulation optimization and nutritional balance monitoring are implemented for low-risk groups.

6. The method as described in claim 5, characterized in that, The functional additives include antioxidants, liver protectants, and microecological regulators to repair the metabolic system and reduce endogenous oxidative stress levels.

7. A sub-health diagnostic system for aquatic animals based on the two-hit theory, characterized in that, include: The risk assessment module is used to calculate the risk value of the first strike and the risk value of the second strike, respectively. The health rating module is used to generate a comprehensive health risk index; The intervention control module is used to implement tiered intervention strategies based on the risk index.

8. The system as described in claim 7, characterized in that, The risk assessment module includes a nutrition monitoring unit and a toxicology monitoring unit. The nutrition monitoring unit is used to collect feed energy composition data and calculate metabolic load, while the toxicology monitoring unit is used to detect the content of non-nutritive toxic substances.

9. The system as described in claim 7, characterized in that, The health rating module uses a multivariate linear model to calculate the health risk index. The input parameters include metabolic load indicators, toxicological stress indicators, and clinical physiological indicators.

10. The system as described in claim 7, characterized in that, The intervention control module automatically generates feed replacement plans, water quality control instructions, and feeding optimization suggestions based on the classification results of the health risk index, realizing integrated control of diagnosis and intervention.

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