Hypoxia-resistant traditional chinese medicine preparation for acclimatizing epinephelus malabaricus and preparation method and application thereof

Abstract

The anti-hypoxia traditional Chinese medicine preparation for domesticating Pomatomus saltatrix, a preparation method thereof and application are prepared from notoginseng, rhodiola, honeysuckle, dandelion, isatis leaf, houttuynia cordata, medlar, licorice, salvia miltiorrhiza, astragalus and acanthopanax, and have the advantages of fast effect, low cost, no toxicity and no drug withdrawal period, can effectively prevent the accumulation of anti-hypoxia agents in the body of Pomatomus saltatrix, causing harm to the human body after eating; the preparation method of the anti-hypoxia traditional Chinese medicine preparation for domesticating Pomatomus saltatrix is simple, has strong repeatability, and is suitable for mass production in industry; the method for ecologically transporting Pomatomus saltatrix by ice temperature, which applies the anti-hypoxia traditional Chinese medicine preparation for domesticating Pomatomus saltatrix to the domestication stage before transportation of the Pomatomus saltatrix, helps to reduce the stress resistance of the Pomatomus saltatrix and reduce the mortality rate of the Pomatomus saltatrix during transportation of the Pomatomus saltatrix.

Description

Technical Field

[0001] This invention relates to the field of traditional Chinese medicine domestication and aquaculture of humpback perch, and in particular to anti-hypoxia traditional Chinese medicine preparations for domesticating humpback perch, their preparation methods and applications. Background Technology

[0002] The humpback perch, also known as the mouse grouper, is priced as high as 700 yuan per kilogram, making it the most expensive species in the grouper subfamily. Its high price can bring considerable profit margins to fish farmers, indicating that its aquaculture prospects are quite optimistic.

[0003] However, its production and habitat are limited, mainly concentrated in tropical waters such as China, Japan, Indonesia, and Australia, resulting in low resource quantities. Furthermore, the humpback bass is aggressive and highly stress-prone, making it extremely prone to death during farming and transportation. Even under oxygenated conditions, humpback bass will exhibit strong stress responses to noise and vibration, such as ramming into the tank, violent jumping, and biting each other. Excessive stress can lead to reduced appetite, stunted growth, weakened immunity, increased susceptibility to disease, and even death. Therefore, humpback bass commonly found in inland markets are typically preserved and transported using refrigeration or freezing. However, refrigeration and freezing reduce the nutritional value of the humpback bass, and the formation of crystal water within the flesh deteriorates the meat quality.

[0004] To meet the demands of fresh transportation, ecological ice-temperature technology is typically used to treat humpback bass in order to reduce mortality during transport.

[0005] Ecological ice-temperature technology is a commonly used, green, healthy, and safe live transport technology. However, humpback bass will experience stress responses due to temperature changes. The key to reducing stress responses lies in anti-hypoxia acclimatization, such as using anti-hypoxia agents to reduce the sensitivity of fish. Commonly used chemical anti-hypoxia agents include MS-222, cloves, and nitrous oxide. However, some of these anti-hypoxia agents have a drug decay period, and some can accumulate in the fish's body, causing irreversible damage to the humpback bass's nervous system. Therefore, there are many safety hazards associated with using the above-mentioned chemical anti-hypoxia agents.

[0006] While traditional Chinese medicine (TCM) anti-hypoxia agents are less prone to accumulation in the body and have less harmful effects on humpback perch, existing TCM preparations involve complex extraction processes and low efficiency. Furthermore, methods for using TCM anti-hypoxia agents to raise and domesticate humpback perch have not been provided. Therefore, it is necessary to develop an anti-hypoxia TCM preparation for domesticating humpback perch, along with its preparation method and application. Summary of the Invention

[0007] In order to solve the technical problem that chemical hypoxia inhibitors used in the process of domesticating humpback bass to resist hypoxia can accumulate in the humpback bass's body and easily cause harm to the human body after consumption, this invention provides a traditional Chinese medicine preparation for domesticating and raising humpback bass to resist hypoxia, which prevents the accumulation of harmful components in the humpback bass's body and has less harm to the human body after consumption.

[0008] Prepared from the following parts by weight of active pharmaceutical ingredient:

[0009]

[0010]

[0011] In this traditional Chinese medicine formula, the principal herbs are Panax notoginseng and Rhodiola rosea; the assistant herbs are Lonicera japonica, Taraxacum mongolicum, Isatis tinctoria leaf, Houttuynia cordata, Lycium barbarum, Crataegus pinnatifida, and Glycyrrhiza uralensis; the adjuvant herbs are Salvia miltiorrhiza and Astragalus membranaceus; and the guiding herb is Acanthopanax senticosus. The individual effects of each of these herbs are as follows:

[0012]

Sanqi

[0013] Rhodiola rosea: Sweet, bitter, and neutral in nature. It enters the lung and heart meridians; it has the effect of invigorating qi and promoting blood circulation; it contains rhodioloside, which can enhance the vitality of hypoxic myocardial cells, improve the hypoxia resistance of humpback perch, and reduce its stress response to changes in oxygen levels.

[0014] When humpback perch experience stress, cortisol levels rise and cellular oxygen levels decrease. Intense struggling exacerbates their anxiety, leading to an overreaction. Using Panax notoginseng and Rhodiola rosea as principal herbs can reduce cortisol levels and increase cellular oxygen levels, thereby mitigating the severity of their overreaction.

[0015] Honeysuckle: Sweet and cold in nature; it has the effects of clearing heat and detoxifying, and dispersing wind-heat.

[0016] Dandelion: bitter, sweet, and cold in nature; it has the effects of clearing heat and detoxifying, reducing swelling and dissipating nodules.

[0017]

Daqingye

[0018] Houttuynia cordata: pungent and slightly cold; it has the effects of clearing heat and detoxifying, eliminating carbuncles and draining pus, and promoting urination.

[0019] Goji berries: sweet and neutral in nature; used for deficiency of essence and blood, and blurred vision.

[0020] Hawthorn: Sour, sweet, and slightly warm; it has the effects of promoting qi circulation, removing blood stasis, clearing turbidity, and lowering lipids.

[0021] Licorice: Sweet and neutral in nature; it has the effects of tonifying the spleen and replenishing qi, and mitigating the harshness of other medications.

[0022]

Salvia miltiorrhiza

[0023] Astragalus membranaceus: Sweet, slightly warm; used for qi deficiency and fatigue, and for tonifying qi and raising yang.

[0024]

Acanthopanax senticosus

[0025] The aforementioned principal herbs are warm in nature. When combined with the slightly cold herbs, the warm nature of the principal herbs is reduced, and the efficacy of the principal herbs is enhanced. Among the adjuvant herbs, Danshen and Huangqi are slightly cold and slightly warm, which work together to promote blood circulation and increase the oxygen content in the cells of the humpback perch, thereby reducing its resistance to environmental changes. The herb Acanthopanax senticosus is rich in antioxidants, which can eliminate free radicals, reduce the damage of oxidative stress to the body, and regulate the body's ability to resist stress, thereby enhancing the body's ability to adapt to various stress factors. This allows the humpback perch to better absorb the principal, adjuvant, and adjuvant herbs, helping the traditional Chinese medicine system to exert its best effects.

[0026] Furthermore, the traditional Chinese medicine preparation for the anti-hypoxic domestication and breeding of humpback perch is prepared from the following raw materials in parts by weight:

[0027]

[0028] Specifically, the traditional Chinese medicine preparation for the anti-hypoxic domestication and breeding of humpback perch is prepared from the following raw materials in parts by weight: 35 parts of Panax notoginseng, 15 parts of Rhodiola rosea, 10 parts of honeysuckle, 4 parts of dandelion, 6 parts of Isatis indigotica, 5 parts of Houttuynia cordata, 6 parts of hawthorn, 6 parts of wolfberry, 8 parts of licorice, 30 parts of Salvia miltiorrhiza, 10 parts of Astragalus membranaceus, and 5 parts of Acanthopanax senticosus.

[0029] More specifically, the Panax notoginseng is produced in Wenshan, Yunnan; the Rhodiola rosea is produced in Tibet; and the honeysuckle, dandelion, and houttuynia cordata are produced in Guangdong. Through experimental comparison, the inventors found that the Chinese medicines produced in the above-mentioned production areas can achieve the best application effects.

[0030] The second objective of this invention is to provide a method for preparing the above-mentioned traditional Chinese medicine preparation, comprising the following steps:

[0031] S1. Preparation of main materials: Pulverize the weighed Panax notoginseng, Rhodiola rosea, honeysuckle, dandelion, Isatis indigotica, Houttuynia cordata, wolfberry, hawthorn, licorice, Salvia miltiorrhiza, Astragalus membranaceus, and Acanthopanax senticosus to 90-120 mesh to obtain mixture A.

[0032] S2. At a temperature of 40-60℃, add 0.2-0.3 parts of the mixed enzyme to 600-800ml of purified water, stir well, and adjust the pH of the mixed enzyme solution to 4-5 with 10% hydrochloric acid to obtain the mixed enzyme extract.

[0033] S3. Add 5-6 times the mass of the mixed enzyme extract to the container, and soak for 8-10 hours at a temperature of 40-60℃.

[0034] S4. Starting from the 6th hour in step S3, collect the filtrate at a uniform flow rate until the time expires and the filtrate stops flowing out, obtaining filtrate B and filter residue C.

[0035] S5. Add 70% ethanol, which is 6-8 times the mass of filter residue C, to the container and the filter residue of S4, and soak for 7-10 hours to obtain drug solution D.

[0036] S6. Mix drug solution D and filtrate B evenly, and concentrate under reduced pressure to obtain a concentrated solution E without alcohol odor. Then, at 45-50℃, adsorb the concentrated solution E using macroporous resin, and elute the macroporous resin with 70% ethanol until it is colorless. Collect the eluent, and then concentrate the eluent under reduced pressure to 1 / 4 to 1 / 5 of the original volume. Add 1 to 2 times the total mass of 30% butanediol to redissolve, filter, and the product is obtained.

[0037] Enzymes exhibit high activity at suitable pH and temperature; and plant cells contain 90% polysaccharides, mainly cellulose, hemicellulose, and pectin. By using enzymatic hydrolysis to break the cell walls of the traditional Chinese medicine before solvent extraction, the extraction concentration can be significantly increased, thereby enhancing the solubility of effective components such as flavonoids and polyphenols in the traditional Chinese medicine preparation of this invention, thus improving the effect of the anti-hypoxia agent. Furthermore, the use of macroporous resin can remove unnecessary impurities, further purifying the preparation and enhancing the anti-hypoxia effect.

[0038] Butanediol can be used as both a solute and a solvent in this invention, thereby enhancing the solubility of traditional Chinese medicine preparations during use.

[0039] Specifically, the mixing ratio of the mixed enzymes is cellulase and pectinase at a mass ratio of 1-1.5:1.

[0040] More specifically, the macroporous resin is AB-8 macroporous resin, which is pretreated for 10-20 minutes at an ultrasonic frequency of 20kHz-100kHz before use. AB-8 is a polystyrene-type weakly polar adsorption resin with certain ester groups on its surface, which improves its hydrophilicity, but the adsorption mechanism is still hydrophobic adsorption. This resin has a large specific surface area and pore size, making it suitable for adsorbing various Chinese herbal medicine components with certain hydrophobicity. It has a large adsorption capacity, is easy to elute, and has good adsorption kinetics. It is stable to heat, organic solvents, and acids and alkalis under normal operating conditions, thus having a long service life. It does not adsorb proteins, sugars, inorganic acids, alkalis, salts, or small molecule hydrophilic organic compounds, thus it can separate common Chinese herbal medicine components from these substances.

[0041] Experiments have shown that pretreating AB-8 macroporous resin with ultrasonic frequencies of 20kHz-100kHz for 10-20 minutes can increase its adsorption capacity for flavonoids.

[0042] The third objective of this invention is to provide a method for ecological ice-temperature transportation of humpback perch after domesticating them using the aforementioned traditional Chinese medicine preparations.

[0043] 48-168 hours before transportation, add the above-mentioned Chinese medicine preparation to the water body for domestication and breeding, and carry out hypoxia resistance domestication of humpback bass under the condition of water temperature of 16-18℃. After domestication is completed, use ecological ice temperature transportation.

[0044] Furthermore, the concentration of the traditional Chinese medicine preparation in the domestication and breeding water is 20 mg / L to 80 mg / L.

[0045] Specifically, the ecological ice-temperature transportation process uses a temperature of 2–14°C for transportation.

[0046] Before transportation, it is necessary to improve the humpback bass's ability to withstand hypoxia, so it needs to be domesticated in advance. The inventors have determined through experimental research that the most suitable period for advance domestication is 48-168 hours.

[0047] Compared with the prior art, the present invention has the following advantages:

[0048] 1. The traditional Chinese medicine preparation for anti-hypoxia domestication of humpback bass provided in this application is made from Panax notoginseng, Rhodiola rosea, honeysuckle, dandelion, Isatis tinctoria leaf, Houttuynia cordata, wolfberry, licorice, Salvia miltiorrhiza, Astragalus membranaceus, and Acanthopanax senticosus. It has the advantages of rapid effect, low cost, no toxicity and no drug decline period. It can effectively prevent the accumulation of anti-hypoxia agents in the humpback bass and prevent harm to the human body after consumption.

[0049] 2. The method for preparing the anti-hypoxia traditional Chinese medicine preparation for domesticating humpback bass provided in this application is simple, highly reproducible, and suitable for mass production in industry.

[0050] 3. This application also provides a method for transporting humpback perch in an ecologically controlled ice-temperature environment. The traditional Chinese medicine preparation for resistant hypoxia-induced domestication of humpback perch is applied to the domestication stage of the humpback perch before transportation, which helps to reduce the stress resistance and mortality of humpback perch during transportation. Attached Figure Description

[0051] Figure 1 This is a graph showing the changes in venous oxygen saturation of domesticated and farmed humpback perch at different concentrations of traditional Chinese medicine preparations at 8±1℃ in Example 6.

[0052] Figure 2 This is a graph showing the changes in venous oxygen saturation of domesticated and farmed humpback perch at different concentrations of traditional Chinese medicine preparations at 18±1℃ in Example 6.

[0053] Figure 3 This is a graph showing the changes in venous oxygen saturation of domesticated humpback perch at different concentrations of traditional Chinese medicine preparations at 28±1℃ in Example 6. Detailed Implementation

[0054] The specific technical solutions of the present invention are described below with reference to specific embodiments 1-7:

[0055] Examples 1-5 specifically illustrate the advantages of the traditional Chinese medicine preparation for acclimating humpback bass and its preparation method in this invention; Examples 6-7 illustrate the advantages of the application of the traditional Chinese medicine preparation for acclimating humpback bass in this invention.

[0056] The origin and specifications of the humpback bass used in the embodiments of this invention are as follows:

[0057] Purchased from Penghai Aquatic Products Trading Co., Ltd., Zhongshan City, Guangdong Province. The original breeding base was the temporary holding area of ​​the Agricultural Products Trading Center in Xiqu District, Zhongshan City, Guangdong Province. The seawater salinity of the breeding area was 2.2%, the pH value was 7.3±0.2, the water flow rate was 5L / min, the temperature was (27±1)℃, and the dissolved oxygen content (DO) was maintained at (7.3±0.5)mg / L. Artificial compound feed was given at 9:00 and 17:00 every day. The humpback bass in the following examples should be of similar size (1260.39±15.60)g, (25.94±0.95)cm, healthy without external injury, and growing well.

[0058] Example 1:

[0059] The following is a total Chinese herbal medicine content of 450g, composed of the following ingredients:

[0060]

[0061]

[0062] Table 1

[0063] Preparation steps:

[0064] S1. Preparation of main materials: Pulverize the weighed Panax notoginseng, Rhodiola rosea, honeysuckle, dandelion, Isatis indigotica, Houttuynia cordata, wolfberry, hawthorn, licorice, Salvia miltiorrhiza, Astragalus membranaceus, and Acanthopanax senticosus to 90 mesh to obtain mixture A1.

[0065] S2. At a temperature of 40℃, add 0.2 parts of the mixed enzyme to 600ml of purified water, stir evenly at 1000r / min, and adjust the pH of the mixed enzyme solution to 4 with 10% hydrochloric acid to obtain the mixed enzyme extract.

[0066] S3. Add a mixed enzyme extract (cellulase and pectinase in a mass ratio of 1:1) to a container at a mass of 5 times the total mass of mixture A1, and soak for 8 hours at a temperature of 40°C.

[0067] S4, starting from the 6th hour in step S3, at 0.1m 3 Collect the filtrate at a flow rate of / s until the filtrate no longer flows out, to obtain filtrate B1 and filter residue C1;

[0068] S5. Add 70% ethanol to the container and the filter residue of S4 in an amount equal to 6 times the mass of filter residue C, and soak for 10 hours to obtain drug solution D1.

[0069] S6. Mix drug solution D and filtrate B evenly, concentrate under reduced pressure until there is no alcohol odor to obtain concentrated solution E. Then, add concentrated solution E1 to AB-8 macroporous resin that has been pretreated at 100kHz for 10 minutes at 45-50℃, elute with 70% ethanol until the macroporous resin is colorless, collect the eluent, concentrate the eluent under reduced pressure to 1 / 4 of the original volume, add 2 times the total mass of 30% butanediol to redissolve, filter, and the product is obtained.

[0070] Example 2:

[0071] The following is a total Chinese herbal medicine content of 450g, composed of the following ingredients:

[0072] Sanqi 35 copies Rhodiola rosea 15 copies honeysuckle 10 copies Dandelion 4 copies Daqingye 6 copies Houttuynia cordata 5 copies Goji berries 6 copies hawthorn 8 copies licorice 8 copies Salvia miltiorrhiza 30 copies Astragalus 10 copies Acanthopanax senticosus 5 copies

[0073] Table 2

[0074] Preparation steps:

[0075] S1. Preparation of main materials: Pulverize the weighed Panax notoginseng, Rhodiola rosea, honeysuckle, dandelion, Isatis tinctoria leaf, houttuynia cordata, wolfberry, hawthorn, licorice, salvia miltiorrhiza, astragalus membranaceus and Acanthopanax senticosus to 120 mesh to obtain mixture A2.

[0076] S2. At a temperature of 45℃, add 0.3 parts of the mixed enzyme to 600-800ml of purified water, stir evenly at 2000r / min, and adjust the pH of the mixed enzyme solution to 4.5 with 10% hydrochloric acid to obtain the mixed enzyme extract.

[0077] S3. In a container, add a mixed enzyme extract (cellulase and pectinase at a mass ratio of 1.5:1) that is 6 times the total mass of mixture A2, and soak for 8 hours at a temperature of 45°C.

[0078] S4, starting from the 6th hour in step S3, at 0.1m 3 Collect the filtrate at a flow rate of / s until the filtrate no longer flows out, and obtain filtrate B2 and filter residue C2.

[0079] S5. Add 70% ethanol to the container and the filter residue of S4 in an amount equal to 6 times the mass of filter residue C2, soak for 7 hours to obtain drug solution D2.

[0080] S6. Mix drug solution D and filtrate B evenly, concentrate under reduced pressure to a boiling point of about 65°C until there is no alcohol odor to obtain concentrated solution E2. Then, add concentrated solution E to AB-8 macroporous resin that has been pretreated at 20kHz for 20 minutes at 45°C, elute with 70% ethanol until the macroporous resin is colorless, collect the eluent, concentrate the eluent under reduced pressure to 1 / 5 of the original volume, add 1 times the total mass of 30% butanediol to redissolve, filter, and the product is obtained.

[0081] Example 3:

[0082] The following is a total Chinese herbal medicine content of 450g, composed of the following ingredients:

[0083]

[0084]

[0085] Table 3

[0086] The remaining conditions in this embodiment are the same as in Embodiment 2.

[0087] Example 4:

[0088] In this embodiment, the enzymatic hydrolysis steps S2-S4 are removed, and the remaining conditions and components are the same as in Example 2.

[0089] Example 5:

[0090] In this embodiment, the AB-8 macroporous resin was not subjected to ultrasonic treatment, and the remaining conditions and components were the same as in Example 2.

[0091] Results Verification: The traditional Chinese medicine preparations for anti-hypoxic domestication and breeding of humpback bass prepared in Examples 1-5 were diluted to a concentration of 60 mg / L and added to a water body with a stocking density of 4 fish / 10L at 18±1℃. The COR (Coefficient of Recovery) and HSP70 levels in the serum of humpback bass were tested and recorded using conventional methods on days 7 and 14. (The significance of these two data points is that COR is an important indicator for evaluating the stress response of fish; a lower COR value indicates less energy loss during swimming, thus indicating lower stress levels. HSP70, or heat shock protein 70, acts as a molecular chaperone in the body, playing a protective role, regulating cellular physiological functions, and improving cellular tolerance to stress responses; its content is positively correlated with stress levels.) The results are shown in Tables 4-5.

[0092]

[0093] Table 4

[0094]

[0095] Table 5

[0096] As can be seen from Tables 4-5, Example 2 is the best embodiment of the present invention. The traditional Chinese medicine preparation prepared using the example has the lowest COR content in the serum of the humpback perch, which is about 111.54 ng / mL, and the HSP70 content in the serum is also at the lowest level, which is about 389.46 pg / mL.

[0097] Furthermore, a comparison between Example 2 and Example 4 reveals that when the enzymatic hydrolysis step is removed, the COR content in the serum of the obtained traditional Chinese medicine preparation and the HSP70 content in the serum are both increased, proving that the enzymatic hydrolysis step helps the present invention achieve the best implementation effect.

[0098] A comparison between Example 2 and Example 5 reveals that, without ultrasonic treatment, the COR content and HSP70 content in the serum of the Chinese herbal preparation obtained from AB-8 macroporous resin were increased, demonstrating that ultrasonic treatment of AB-8 macroporous resin helps improve the anti-hypoxia effect of the anti-hypoxia agent.

[0099] Example 6

[0100] In this embodiment, the optimal concentration of the aforementioned anti-hypoxia agent is determined through simulation. The effects of different concentrations of traditional Chinese medicine anti-hypoxia agents and acclimatized aquaculture water temperatures on humpback bass are investigated. The specific methods are as follows:

[0101] Orthogonal domestication and aquaculture experiments were conducted to verify the concentration of the traditional Chinese medicine preparations and the aquaculture water temperature in this invention.

[0102] The traditional Chinese medicine preparations in this embodiment were prepared in Example 2 and used in the acclimatization test at concentrations of 20 mg / L, 40 mg / L, 60 mg / L, and 80 mg / L, respectively.

[0103] The water temperatures used for domestication and aquaculture in this invention are 8±1℃, 18±1℃, and 28.0±1℃.

[0104] The domestication and rearing density was 4 fish / 10L, and the domestication period was 7 days. During the domestication and rearing process, the oxygen supply was adjusted so that the dissolved oxygen (DO) was gradually reduced from (8±0.5) mg / L to (1±0.5) mg / L at a rate of 1 mg / L per day.

[0105] A control group was established with a temporary holding density of 4 fish / 10L. During the 7-day temporary holding period, no traditional Chinese medicine anti-hypoxia agent was added, and the dissolved oxygen (DO) level was maintained at (8±0.5) mg / L. During the domestication and breeding period, blood was drawn from the tail vein of the humpback bass daily before reducing DO using a 2mL syringe, and the venous blood oxygen saturation was measured.

[0106] Blood samples were collected from the gill vein of humpback bass before and during acclimatization and rearing, every 24 hours. The samples were rapidly centrifuged at 2000 rpm for 10.0 min using a high-speed centrifuge. After separation, the supernatant was collected in test tubes, and serum total protein content was measured. EDTA-K2 anticoagulant tubes were then inserted, and changes in venous oxygen saturation, complete blood count, and biochemical indicators were measured. Spinal venous blood samples were collected from humpback bass before, during, and after acclimatization and rearing. Laboratory sampling was commissioned to Meikang Biotechnology Co., Ltd. for testing.

[0107] The humpback bass were pretreated with traditional Chinese medicine to combat hypoxia and then cooled to their ecological temperate zone dormancy at a rate of 5.0°C / h. The survival rate of the humpback bass was studied.

[0108] All experiments were repeated three times, and the trend of venous oxygen saturation in humpback bass was measured using standard methods (e.g., Figure 1-3 The following data were recorded: blood cell counts of humpback bass raised under hypoxia resistance at 8±1℃ (Table 6), blood cell counts of humpback bass raised under hypoxia resistance at 18±1℃ (Table 7), blood cell counts of humpback bass raised under hypoxia resistance at 28.0±1℃ (Table 8), serum biochemical indicators of humpback bass raised under hypoxia resistance at 8±1℃ (Table 9), serum biochemical indicators of humpback bass raised under hypoxia resistance at 18.0±1℃ (Table 10), and serum biochemical indicators of humpback bass raised under hypoxia resistance at 28.0±1℃ (Table 11). The experimental data were recorded as mean ± standard deviation.

[0109]

[0110] Table 6

[0111]

[0112]

[0113] Table 7

[0114]

[0115]

[0116] Table 8

[0117]

[0118]

[0119] Table 9

[0120]

[0121]

[0122] Table 10

[0123]

[0124] Table 11

[0125] Example 7

[0126] This embodiment investigates the effects of different domestication and rearing temperatures, transport ecological ice temperatures, and transport densities on the survival rate of humpback bass. The critical temperature for humpback bass was determined using the following method:

[0127] Five humpback bass were placed in a plastic tank filled with clean seawater. Homemade ice cubes (wrapped in sealed bags) were used as a cooling source, and the bass were slowly cooled according to the cooling rates in Table 12. During this process, the respiratory rate and activity of the humpback bass at different temperatures were observed and recorded. Based on Table 13, the critical temperature for the humpback bass was defined as the temperature at which extremely irregular breathing began, or even when gill respiration was no longer observable, but the fish were not yet dead.

[0128]

[0129] Table 12

[0130]

[0131]

[0132] Table 13

[0133] After being domesticated and cultured with traditional Chinese medicine anti-hypoxia agents, humpback bass were cooled to the critical temperature according to the gradient cooling pattern in Table 12 before simulated transportation. The simulated transportation experiment included four transportation densities: low-density (2 fish / 10L), medium-density (4 fish / 10L), medium-high density (6 fish / 10L), and high-density (8 fish / 10L), and four transportation temperatures (2℃, 6℃, 10℃, and 14℃). A control group was also included with four transportation densities, with three replicates for each density. The survival rate of humpback bass under different domestication concentrations and transportation densities was monitored at 0, 2, 4, 6, and 8 hours during the simulated transportation period. (The survival time was considered successful when the humpback bass stopped moving their gills and were placed in an oxygenated tank to awaken to stage R5 of the hypoxia resistance and awakening level classification in Table 13, and this time was considered the longest survival time at that domestication concentration, transportation density, and temperature.)

[0134] The survival rates of humpback perch raised under hypoxia resistance during ecological ice-temperature transportation at 8±1℃ are shown in Table 14, 18±1℃, and 28±1℃ respectively.

[0135]

[0136] Table 14

[0137]

[0138]

[0139] Table 15

[0140]

[0141]

[0142] Table 16

[0143] As can be seen from Examples 6-7, in terms of blood cell count and biochemical indicators, the blood indicators of humpback perch were optimal after acclimatization to hypoxia at 18℃ and a concentration of 60 mg / L anti-hypoxia agent. Even during ecological ice-temperature transportation, the survival rate under acclimatization to hypoxia conditions of 8℃ and 18℃ with a concentration of 60 mg / L could reach 100%. However, considering factors such as the amount of anti-hypoxia agent used and energy conservation during ecological ice-temperature transportation, the optimal conditions are 14℃ ecological ice-temperature transportation after acclimatization to hypoxia at 18℃ and a concentration of 60 mg / L anti-hypoxia agent. These are the optimal conditions for the application of the traditional Chinese medicine preparation of this invention for acclimatization to hypoxia in humpback perch.

[0144] The above description is merely a preferred embodiment of this application and is not intended to limit this application. Any modifications, equivalent substitutions, and improvements made within the spirit and principles of this application should be included within the protection scope of this application.

Claims

1. A traditional Chinese medicine preparation for combating hypoxia used in the domestication of humpback perch, characterized in that, It is prepared from the following raw materials in parts by weight, with a total Chinese herbal medicine content of 450g: The method for preparing the traditional Chinese medicine preparation includes the following steps: S1. Preparation of main materials: Pulverize the weighed Panax notoginseng, Rhodiola rosea, honeysuckle, dandelion, Isatis indigotica, Houttuynia cordata, wolfberry, hawthorn, licorice, Salvia miltiorrhiza, Astragalus membranaceus, and Acanthopanax senticosus to 90-120 mesh to obtain mixture A. S2. At a temperature of 40-60℃, add 0.2-0.3 parts of the mixed enzyme to 600-800ml of purified water, stir well, and adjust the pH of the mixed enzyme solution to 4-5 with 10% hydrochloric acid to obtain the mixed enzyme extract. S3. Add 5-6 times the mass of the mixed enzyme extract to the container, and soak for 8-10 hours at a temperature of 40-60℃. S4. Starting from the 6th hour in step S3, collect the filtrate at a uniform flow rate until the time expires and the filtrate stops flowing out, obtaining filtrate B and filter residue C. S5. Add 70% ethanol to the container and the filter residue of S4 in an amount equal to 6-8 times the mass of filter residue C, and soak for 7-10 hours to obtain drug solution D. S6. Mix the medicinal solution D and the filtrate B evenly, and concentrate under reduced pressure to obtain a concentrated solution E without alcohol odor. Then, at 45-50℃, adsorb the concentrated solution E using macroporous resin, and elute the macroporous resin with 70% ethanol until it is colorless. Collect the eluent, and then concentrate the eluent under reduced pressure to 1 / 4 to 1 / 5 of the original volume. Add 1 to 2 times the total mass of 30% butanediol to redissolve, filter, and obtain the traditional Chinese medicine preparation. The macroporous resin is AB-8 macroporous resin, which is pretreated for 10-20 minutes at an ultrasonic frequency of 20kHz-100kHz before use. The mixing ratio of the mixed enzymes is cellulase and pectinase at a mass ratio of 1-1.5:

1.

2. The traditional Chinese medicine preparation according to claim 1, characterized in that, Prepared from the following parts by weight of active pharmaceutical ingredient:

3. The traditional Chinese medicine preparation according to claim 2, characterized in that, It is prepared from the following raw materials in parts by weight: Panax notoginseng 35 parts, Rhodiola rosea 15 parts, Lonicera japonica 10 parts, Taraxacum mongolicum 4 parts, Isatis tinctoria leaf 6 parts, Houttuynia cordata 5 parts, Crataegus pinnatifida 6 parts, Lycium barbarum 6 parts, Glycyrrhiza uralensis 8 parts, Salvia miltiorrhiza 30 parts, Astragalus membranaceus 10 parts, Acanthopanax senticosus 5 parts.

4. The traditional Chinese medicine preparation according to claim 2 or 3, characterized in that: The Panax notoginseng is produced in Wenshan, Yunnan; the Rhodiola rosea is produced in Tibet; and the honeysuckle, dandelion, and houttuynia cordata are produced in Guangdong.

Images