Method for improving fish muscle quality by regulating the intestinal microenvironment of the neuroendocrine system
By adding Lactobacillus bakerii MN047 and lotus root extract to the fish holding tank, the intestinal microbiota was regulated, which solved the problems of neuroendocrine system disorders and muscle quality deterioration caused by fish stress, and achieved muscle quality improvement.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- FARM PROD PROCESSING & NUCLEAR AGRI TECH INST HUBEI ACAD OF AGRI SCI
- Filing Date
- 2024-12-31
- Publication Date
- 2026-05-19
AI Technical Summary
Fish may experience neuroendocrine system disorders and gut microbiota homeostasis imbalances due to stress responses during the fish industry chain, which in turn leads to deterioration in muscle quality.
By adding a mixture of Lactobacillus bakerii MN047 and lotus root extract to the temporary holding tank, the intestinal microbiota was regulated, the secretion of neurotransmitters was reduced, the intestinal microbiota homeostasis was maintained, and muscle quality was improved.
It effectively alleviates the deterioration of muscle quality caused by stress response, improves the firmness and water retention capacity of fish meat, reduces oxidative inflammation response, and enhances the economic value of fish meat.
Smart Images
Figure CN119769445B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of muscle quality regulation of aquatic products, and mainly to a method for improving muscle quality by regulating the intestinal microenvironment through the neuroendocrine system. Background Technology
[0002] In the fish industry chain, many crucial stages can induce stress, such as aquaculture, fishing, transportation, and slaughter, leading to physiological responses, oxidative damage, and metabolic disorders in fish, ultimately resulting in deterioration of muscle quality. The stress response originates from the activation of the central nervous system and the rapidly acting sympathetic-adrenal-medullaecine system. This system releases norepinephrine from the postganglionic sympathetic nerves, adrenaline from the adrenal medulla, and, more slowly, the hypothalamic-pituitary-adrenal axis, ultimately releasing cortisol from the adrenal cortex. The release of cortisol triggers a rapid response in the blood glucose metabolism system and simultaneously activates the GR receptors in the body's immune system, affecting gut microbiota homeostasis. There is a close and bidirectional communication between the central nervous system and the gut and its microbiota mediated by the nervous, endocrine, and immune systems. The gut microbiota and bacterial metabolites are related to the activation of sympathetic and parasympathetic enteric neurons, immune cell activity, and the regulation of neurotransmitter and gut hormone production. The gut microbiota and skeletal muscle also interact. Dysbiosis of the gut microbiota may increase oxidative inflammation, affecting the homeostasis of energy synthesis and metabolism in mitochondria and muscle, ultimately leading to changes in muscle quality. Fish meat firmness and water-holding capacity are important indicators of muscle quality. Shear force represents the good maintenance of fish meat firmness, while pressure loss rate represents water-holding capacity. Decreased firmness and reduced water-holding capacity ultimately result in loose, dull fish meat, reducing its economic value. Fish have a weak ability to alleviate stress; stress at any stage of the supply chain can easily cause neuroendocrine system disorders, gut microbiota homeostasis imbalances, muscle quality deterioration, and even fish death. Therefore, this invention provides a method from the perspective of neurotransmitters-gut microbiota-muscle quality, providing a method to alleviate muscle quality deterioration caused by stress responses by rapidly secreting neurotransmitters to alter the gut microbiota. Summary of the Invention
[0003] The purpose of this invention is to provide a method for improving the muscle quality of fish by regulating the intestinal microenvironment through the neuroendocrine system. This method can alleviate the stress response in fish after stress, secrete neurotransmitters, regulate the intestinal microbiota, and thus alleviate the deterioration of muscle quality caused by stress response.
[0004] To achieve the above objectives, the technical solution adopted by the present invention is: a method for improving fish muscle quality by regulating the intestinal microenvironment through the neuroendocrine system, characterized by comprising the following steps:
[0005] 1) After sterilizing the holding tank, fill it with aerated water to 2 / 3 of its height (the height of the holding tank is 0.5-5m). Use an aerator to adjust the dissolved oxygen in the water to maintain it at 5.0-8.0 mg / L, the water temperature at 16-24℃, the pH at 7.2-8.0, and the ammonia nitrogen content below 0.5 mg / L.
[0006] 2) Add a mixture of 50-100 mg / L Lactobacillus bakerii MN047 and 20-50 mg / L lotus root extract to water and stir to disperse it evenly.
[0007] 3) The stocking density is 20-30 kg / m² 3 Release live freshwater fish into the temporary holding tank and withhold food from the fish during the holding period;
[0008] 4) The temporary holding period is 20-22 days (21 days is the best, but should be adjusted according to the condition of the fish). During the temporary holding period, 2 / 3 of the water should be changed every day. After that, the mixed feed should be added again and the concentration of the mixed feed should be adjusted to be the same as in step 2).
[0009] 5) After the temporary holding period ends, the live freshwater fish are placed in a bucket containing 50 mg / L of anesthetic. After confirming that the live freshwater fish are completely anesthetized, their brain, midgut and muscle tissues are removed, and they are quickly packaged and flash-frozen in liquid nitrogen. They are then placed in a -80 ℃ freezer for testing.
[0010] 6) Neurotransmitters in brain tissue were separated and determined using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry in series. The microbial community in the midgut was identified using 16s microbial diversity analysis. The pressure loss rate of fish meat was determined using a YYW-2 strain-controlled unconfined pressure tester, and the shear force of fish meat was determined using a C-LM3B tenderness tester.
[0011] 7) Analyze the data obtained in step 6) to determine the regulatory effects of the mixed input on neurotransmitters, gut microbiota and muscle quality of live freshwater fish; use correlation analysis to determine the correlation between neurotransmitters, gut microbiota and muscle quality based on the correlation coefficient.
[0012] Furthermore, the temporary holding tank (or suspended water tank) mentioned in step 1) is a circular canvas tank with a diameter of 8 m, a height of 1.5 m, and a water level of 1 m.
[0013] Further, step 1) involves sterilizing the temporary holding tank by using 1 / 10000 volume of methylene blue.
[0014] Furthermore, in step 1), the dissolved oxygen in the water is maintained at 7.0-8.0 mg / L, the water temperature at 16-24 ℃, and the pH at 7.8-8.0.
[0015] Furthermore, in step 1), the ambient temperature is 17℃-24℃.
[0016] Further, in step 2), a mixture of 50-100 mg / L of Lactobacillus bakerii MN047 and 20-50 mg / L of lotus root extract is added to the temporary holding tank, which needs to be fully dissolved in the water.
[0017] Furthermore, in step 2), the Lactobacillus bakerii MN047 strain was screened from homemade mare's milk wine in Xinjiang Uygur Autonomous Region, my country. This strain is capable of producing a variety of broad-spectrum bacteriocins. The antibacterial metabolites of this strain can inhibit foodborne pathogens and regulate the intestinal flora.
[0018] Further, the lotus root extract in step 2) is a mixture containing a certain amount of plant polyphenols extracted from lotus root nodes. The extraction method is as follows: after cutting the lotus root nodes into pieces and grinding them into a paste, it is extracted for 90 minutes at 62 ℃ and pH 4 using 0.15 mg / mL cellulase (50,000 U / g) + pectinase (30,000 U / g) (the mass ratio of cellulase to pectinase is 1:1). After centrifugation at 4000 xg for 20 minutes, the supernatant is collected as the crude extract. Subsequently, it is subjected to preliminary alcohol precipitation with 60% ethanol and then purified with AB-8 macroporous resin. After freeze-drying, the total phenol concentration is measured to be 68.73 mg / 100 mg, thus obtaining the lotus root extract.
[0019] Furthermore, the freshwater fish mentioned in step 3) are healthy, well-formed, and undamaged freshwater fish such as yellow catfish, rainbow trout, mandarin fish, and largemouth bass.
[0020] Furthermore, the fresh freshwater fish mentioned in step 3) are all edible adult fish, with the following weights: yellow catfish 300 ± 10 g, rainbow trout 2000 ± 115 g, mandarin fish 500 ± 50 g, and largemouth bass 400 ± 20 g.
[0021] Furthermore, in step 3), each temporary holding pond is strictly maintained at a stocking density of 20-30 kg / m³. 3 Control the number of fish introduced.
[0022] Furthermore, the concentration of the mixed input of Lactobacillus bakerella and polyphenol extract in step 4) is the same as that in step 2).
[0023] Further, the anesthetic agent described in step 5) has the molecular formula of ethyl m-aminobenzoate methanesulfonate (MS-222), with the molecular formula: C 10 H 15 NO5S.
[0024] Further, the specific method for determining the pressure loss rate in step 6) is as follows: Cut a 4 cm × 4 cm piece of gauze and weigh it (M0). Weigh 2 ± 0.02 g of fresh freshwater fish meat (e.g., rainbow trout) and wrap it in gauze, then weigh it (M1). Place 8 sheets of filter paper of the same size on the top and bottom of the sample. Place the sample wrapped with fish meat in the middle of the YYW-2 type pressure plate and adjust the pressure instrument to 145 Pa and maintain it on the sample for 5 min. After the test, weigh the pressurized gauze and the sample and record the weight (M2). The pressure loss rate of the fish fillet is calculated using the following formula:
[0025]
[0026] Where: X — the pressure loss rate of the sample, %
[0027] M0 — Mass of gauze, g;
[0028] M1—Total mass of gauze and sample before pressure application, g;
[0029] M2 — Total mass of the gauze and sample after pressure application, in g.
[0030] Further, the specific method for measuring shear force in step 6) is as follows: cut the back muscles of the same location into strips of similar shape and size. Before measurement, zero the instrument and cut the blade downwards at a uniform speed until the muscle is completely cut off.
[0031] The beneficial effects of this invention are:
[0032] It effectively reduces the release of excitatory neurotransmitters in the brain caused by stress, while maintaining the homeostasis of the gut microbiota in live freshwater fish, improving the proportion of beneficial microorganisms in the microbiota, and alleviating the deterioration of muscle quality in live freshwater fish caused by stress.
[0033] The present invention uses aerated tap water to purify the body of freshwater fish. Without feeding them, the fish will excrete feces and other harmful substances. In addition, a certain proportion of Lactobacillus bakerii MN047 and lotus root extract are added to reduce the level of neurotransmitters in the brain, maintain the balance of intestinal microorganisms in freshwater fish, and improve the deterioration of muscle quality.
[0034] The antibacterial metabolites of Lactobacillus bakerella MN047 can inhibit foodborne pathogens and regulate intestinal flora. Lotus root extract has antioxidant and antibacterial effects. The combined use of the two can reduce the formation of oxygen free radicals in fish, improve the quality of freshwater fish meat, and reduce the loss of nutrients.
[0035] In summary, the combined use of Lactobacillus bakerii MN047 and lotus root extract provided in this invention can reduce neurotransmitter secretion, maintain intestinal flora homeostasis, and alleviate muscle quality deterioration. Attached Figure Description
[0036] Figure 1 This is a relative level diagram of the intestinal flora contained in the midgut of the fish at the genus level according to the present invention.
[0037] Figure 2a This is a heatmap showing the correlation between fish neurotransmitters, gut microbiota, and muscle quality under low levels of additives (50 mg / L Lactobacillus bakerii MN047 and 20 mg / L lotus root extract) in Example 2 of the present invention.
[0038] Figure 2b This is a heatmap showing the correlation between fish neurotransmitters, gut microbiota, and muscle quality under high levels of additives (100 mg / L Lactobacillus bakerii MN047 and 50 mg / L lotus root extract) in Example 3 of the present invention.
[0039] Table 1 is a statistical table of neurotransmitters with significantly different expression levels in the fish brain of the present invention.
[0040] Table 2 shows the fish muscle quality evaluation indicators of this invention. Detailed Implementation
[0041] In Examples 2-3 below, the lotus root extract is a mixture containing a certain amount of plant polyphenols extracted from lotus root nodes. The extraction method is as follows: the lotus root nodes are cut into pieces and ground into a paste. Then, the extract is prepared by using 0.15 mg / mL cellulase (50,000 U / g) + pectinase (30,000 U / g) (the mass ratio of cellulase to pectinase is 1:1) at a temperature of 62 ℃ and pH 4 for 90 minutes. After centrifugation at 4000 xg for 20 minutes, the supernatant is collected as the crude extract. Subsequently, the crude extract is subjected to preliminary alcohol precipitation with 60% ethanol and then purified with AB-8 macroporous resin. After freeze-drying, the total phenol concentration is measured to be 68.73 mg / 100 mg, thus obtaining the lotus root extract.
[0042] In Examples 1-3 below, the anesthetic has the molecular formula of ethyl m-aminobenzoate methanesulfonate (MS-222), with the molecular formula: C 10 H 15 NO5S.
[0043] Example 1 (Control Group Example)
[0044] A method for improving fish muscle quality by regulating the gut microenvironment using the neuroendocrine system includes the following steps:
[0045] 1) Sterilize the holding tank before use: Sterilize the holding tank with 1 / 10000 volume of methylene blue;
[0046] After sterilizing the holding tank, fill it with clean water (the holding tank is 8 m in diameter and 1.5 m high, with a stocking density of approximately 30 kg / m³). 3 The circular canvas pool is used to control the water depth at 1 m. Before the live fish are put into the holding pool, the water in the pool is aerated for one day. The holding pool is oxygenated with the provided aerator to ensure that the dissolved oxygen in the water is 7.0-8.0 mg / L, the water temperature is 16-24℃, and the pH is 7.8-8.0.
[0047] 2) Randomly select 50 healthy, well-formed, and undamaged live largemouth bass weighing 400 ± 20 g and place them in a temporary holding tank. Adjust the stocking density to 30 kg / m². 3 No substances were added; this group was designated as the control group.
[0048] 3) The temporary holding period is adjusted to 21 days. During the temporary holding period, the water will be changed once a day, and the amount of water changed will be 2 / 3 of the original water volume.
[0049] 4) Fasting will be enforced throughout the entire temporary care period;
[0050] 5) After the temporary holding period ends, the live largemouth bass are placed in a bucket containing 50 mg / L of anesthetic. After confirming that the live freshwater fish are completely anesthetized, professionals remove their brain, midgut, and muscle tissues, which are then quickly packaged and flash-frozen in liquid nitrogen and then placed in a -80 ℃ freezer for testing.
[0051] 6) Neurotransmitters in brain tissue were separated and determined using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry in series. The microbial community in the midgut was identified using 16s microbial diversity analysis. The pressure loss rate of fish meat was determined using a YYW-2 strain-controlled unconfined pressure tester, and the shear force of fish meat was determined using a C-LM3B tenderness tester.
[0052] 7) Analyze the data obtained in 6) to determine the regulatory effects on neurotransmitters, gut microbiota and muscle quality of live freshwater fish; use correlation analysis to determine the correlation between neurotransmitters, gut microbiota and muscle quality based on the correlation coefficient.
[0053] Example 2
[0054] A method for improving fish muscle quality by regulating the gut microenvironment using the neuroendocrine system includes the following steps:
[0055] 1) Sterilize the holding tank before use: Sterilize the holding tank with 1 / 10000 volume of methylene blue;
[0056] After sterilizing the holding tank, aerated water is poured into it (the holding tank is 8 m in diameter and 1.5 m high, with a stocking density of approximately 30 kg / m²). 3 The circular canvas pool is used to control the water depth at 1 m. Before the live fish are put into the holding pool, the water in the pool is aerated for one day. The holding pool is oxygenated with the provided aerator to ensure that the dissolved oxygen in the water is 7.0-8.0 mg / L, the water temperature is 16-24 ℃, and the pH is 7.8-8.0.
[0057] 2) Add a mixture of 50 mg / L Lactobacillus bakerii MN047 and 20 mg / L lotus root extract to water and stir to disperse it evenly.
[0058] 3) Randomly select 50 healthy, well-formed, and undamaged live largemouth bass weighing 400 ± 20 g and place them in a temporary holding tank. Adjust the stocking density to 30 kg / m². 3 During the temporary holding period, the fish were fasted; this group was designated as experimental group 1.
[0059] 4) The temporary holding period is adjusted to 21 days. During the temporary holding period, the water is changed once a day, and the amount of water changed is 2 / 3 of the original water volume. After that, the mixed feed from step 2) is added again, and the concentration of the mixed feed is adjusted to be consistent with that in step 2). Feeding is prohibited throughout the entire temporary holding period.
[0060] 5) After the temporary holding period ends, the live largemouth bass are placed in a bucket containing 50 mg / L of anesthetic. After confirming that the live freshwater fish are completely anesthetized, professionals remove their brain, midgut, and muscle tissues, which are then quickly packaged and flash-frozen in liquid nitrogen and then placed in a -80 ℃ freezer for testing.
[0061] 6) Neurotransmitters in brain tissue were separated and determined using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry in series. The microbial community in the midgut was identified using 16s microbial diversity analysis. The pressure loss rate of fish meat was determined using a YYW-2 strain-controlled unconfined pressure tester, and the shear force of fish meat was determined using a C-LM3B tenderness tester.
[0062] 7) Analyze the data obtained in step 6) to determine the regulatory effects of the mixed input on neurotransmitters, gut microbiota and muscle quality of live freshwater fish; use correlation analysis to determine the correlation between neurotransmitters, gut microbiota and muscle quality based on the correlation coefficient.
[0063] Example 3
[0064] A method for improving fish muscle quality by regulating the gut microenvironment using the neuroendocrine system includes the following steps:
[0065] 1) Sterilize the holding tank before use: Sterilize the holding tank with 1 / 10000 volume of methylene blue;
[0066] After sterilizing the holding tank, aerated water is poured into it (the holding tank is 8 m in diameter and 1.5 m high, with a stocking density of approximately 30 kg / m²). 3 The circular canvas pool is used to control the water depth at 1 m. Before the live fish are put into the holding pool, the water in the pool is aerated for one day. The holding pool is oxygenated with the provided aerator to ensure that the dissolved oxygen in the water is 7.0-8.0 mg / L, the water temperature is 16-24 ℃, and the pH is 7.8-8.0.
[0067] 2) Add a mixture of 100 mg / L Lactobacillus bakerii MN047 and 50 mg / L lotus root extract to water and stir to disperse it evenly.
[0068] 3) Randomly select 50 healthy, well-formed, and undamaged live largemouth bass weighing 400 ± 20 g and place them in a temporary holding tank. Adjust the stocking density to 30 kg / m². 3 During the temporary holding period, the fish were fasted; this group was designated as experimental group 2.
[0069] 4) The temporary holding period is adjusted to 21 days. During the temporary holding period, the water is changed once a day, and the amount of water changed is 2 / 3 of the original water volume. After that, the mixed feed from step 2) is added again, and the concentration of the mixed feed is adjusted to be consistent with that in step 2). Feeding is prohibited throughout the entire temporary holding period.
[0070] 5) After the temporary holding period ends, the live largemouth bass are placed in a bucket containing 50 mg / L of anesthetic. After confirming that the live freshwater fish are completely anesthetized, professionals remove their brain, midgut, and muscle tissues, which are then quickly packaged and flash-frozen in liquid nitrogen and then placed in a -80 ℃ freezer for testing.
[0071] 6) Neurotransmitters in brain tissue were separated and determined using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry in series. The microbial community in the midgut was identified using 16s microbial diversity analysis. The pressure loss rate of fish meat was determined using a YYW-2 strain-controlled unconfined pressure tester, and the shear force of fish meat was determined using a C-LM3B tenderness tester.
[0072] 7) Analyze the data obtained in step 6) to determine the regulatory effects of the mixed input on neurotransmitters, gut microbiota and muscle quality of live freshwater fish; use correlation analysis to determine the correlation between neurotransmitters, gut microbiota and muscle quality based on the correlation coefficient.
[0073] In this embodiment, in the preliminary prediction of fresh fish muscle quality based on respiratory rate, the user only needs to perform the operation according to steps 1)-4), without repeating the following steps.
[0074] Referring to Table 1, the present invention can effectively reduce the secretion of excitatory neurotransmitters histamine and kynurenine by adding 50 mg / L of Lactobacillus bakerii MN047 and 20 mg / L of lotus root extract or 100 mg / L of Lactobacillus bakerii MN047 and 50 mg / L of lotus root extract, thereby reducing the stress response to the nervous, endocrine and immune systems of fish.
[0075] Reference Figure 1 The present invention can effectively maintain intestinal microbial homeostasis by adding 50 mg / L of Lactobacillus bakerii MN047 and 20 mg / L of lotus root extract or 100 mg / L of Lactobacillus bakerii MN047 and 50 mg / L of lotus root extract, thereby reducing the number of pathogenic bacteria in the intestine of largemouth bass and increasing the abundance of beneficial bacteria in the intestine.
[0076] Referring to Table 2, the present invention can effectively improve muscle quality by adding 50 mg / L of Lactobacillus bakerii MN047 and 20 mg / L of lotus root extract or 100 mg / L of Lactobacillus bakerii MN047 and 50 mg / L of lotus root extract.
[0077] Reference Figure 2a , Figure 2b The present invention can reduce neurotransmitter secretion, maintain intestinal flora homeostasis, and alleviate muscle quality deterioration by adding 50 mg / L of Lactobacillus baker MN047 and 20 mg / L of lotus root extract or 100 mg / L of Lactobacillus baker MN047 and 50 mg / L of lotus root extract.
[0078] Table 1 shows the statistical table of neurotransmitters with significantly different expression levels in the fish brain of this invention.
[0079]
[0080] Table 2 shows the fish muscle quality evaluation indicators of this invention.
[0081]
[0082] The upper and lower limits of the dosage of Lactobacillus bakerella and lotus root extract mentioned in this invention can achieve the present invention. The yellow catfish, rainbow trout, mandarin fish, etc. can all achieve the present invention. The upper and lower limits of dissolved oxygen, water temperature, and pH value can also achieve the present invention. Examples are not listed here.
Claims
1. A method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system, characterized in that... Includes the following steps: 1) After sterilizing the holding tank, fill it with aerated water to 2 / 3 of its height. Use an aerator to adjust the dissolved oxygen in the water to maintain it at 5.0 - 8.0 mg / L, the water temperature at 16 - 24 ℃, the pH at 7.2 - 8.0, and the ammonia nitrogen content below 0.5 mg / L. 2) Add a mixture of 50-100 mg / L Lactobacillus bakerii MN047 and 20-50 mg / L lotus root extract to water and stir to disperse it evenly. The lotus root extract is a mixture containing a certain amount of plant polyphenols extracted from lotus root nodes. The extraction method is as follows: the lotus root nodes are cut into pieces and ground into a paste. Then, 0.15 mg / mL cellulase + pectinase (mass ratio of cellulase to pectinase 1:1) is used for extraction at 62 ℃ and pH 4 for 90 minutes. After centrifugation at 4000 xg for 20 minutes, the supernatant is collected as the crude extract. Subsequently, preliminary alcohol precipitation is performed with 60% ethanol, followed by purification with AB-8 macroporous resin. After freeze-drying, the total phenol concentration is measured to be 68.73 mg / 100 mg, thus obtaining the lotus root extract. 3) The stocking density is 20-30 kg / m² 3 Release live freshwater fish into the temporary holding tank and withhold food from the fish during the holding period; 4) The temporary holding period is 20-22 days. During the temporary holding period, 2 / 3 of the water volume needs to be changed every day. After that, the mixed feed should be added again and the concentration of the mixed feed should be adjusted to be consistent with step 2). 5) After the temporary holding period ends, the live freshwater fish are placed in a bucket containing 50 mg / L of anesthetic. After confirming that the live freshwater fish are completely anesthetized, their brain, midgut and muscle tissue are removed, and they are quickly packaged and flash-frozen in liquid nitrogen. They are then placed in a -80 ℃ freezer for testing. 6) Neurotransmitters in brain tissue were separated and determined using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry in series. The microbial community in the midgut was identified using 16s microbial diversity analysis. The pressure loss rate of fish meat was determined using a YYW-2 strain-controlled unconfined pressure tester, and the shear force of fish meat was determined using a C-LM3B tenderness tester. 7) Analyze the data obtained in step 6) to determine the regulatory effects of the mixed input on neurotransmitters, gut microbiota and muscle quality of live freshwater fish; use correlation analysis to determine the correlation between neurotransmitters, gut microbiota and muscle quality based on the correlation coefficient.
2. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: The temporary holding tank mentioned in step 1) is a circular canvas tank with a diameter of 8 m, a height of 1.5 m, and a water level of 1 m. The sterilization treatment of the temporary holding tank in step 1) is to use 1 / 10000 volume of methylene blue to sterilize the temporary holding tank.
3. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: In step 1), ensure that the dissolved oxygen in the water is 7.0-8.0 mg / L, the water temperature is 16-24 ℃, and the pH is 7.8-8.
0.
4. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: Step 3) The fresh freshwater fish mentioned are healthy, well-formed, and undamaged yellow catfish, rainbow trout, mandarin fish, and largemouth bass.
5. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: Step 3) The fresh freshwater fish mentioned are all edible adult fish, with the following weights: yellow catfish 300 ± 10 g, rainbow trout 2000 ± 115 g, mandarin fish 500 ± 50 g, and largemouth bass 400 ± 20 g.
6. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: Step 5) The anesthetic has the molecular formula of ethyl m-aminobenzoate methanesulfonate, molecular formula: C 10 H 15 NO5S.
7. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: Step 6) The specific method for determining the pressure loss rate is as follows: Cut a 4 cm × 4 cm piece of gauze and weigh it. Weigh 2 ± 0.02 g of fresh freshwater fish meat sample, wrap it in gauze, and weigh it. Place 8 sheets of filter paper of the same size on the top and bottom of the sample. Place the sample wrapped with fish meat in the middle of the YYW-2 type pressure plate and adjust the pressure instrument to 145 Pa and maintain it on the sample for 5 min. After the test, weigh the pressurized gauze and the sample and record it as M2. The pressure loss rate of the fish fillet is calculated using the following formula: Where: X — the pressure loss rate of the sample, % M0 — Mass of gauze, g; M1—Total mass of gauze and sample before pressure application, g; M2 — Total mass of the gauze and sample after pressure application, in g.
8. The method for improving fish muscle quality by regulating the intestinal microenvironment using the neuroendocrine system according to claim 1, characterized in that: Step 6) The specific method for measuring shear force is as follows: cut the back muscles of the same location into strips of similar shape and size. Before measurement, zero the instrument and cut the blade downwards at a constant speed until the muscle is completely cut off.