CN113475452A - Method for compound keeping-alive of slow-release anesthetic - Google Patents
Method for compound keeping-alive of slow-release anesthetic Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K63/00—Receptacles for live fish, e.g. aquaria; Terraria
- A01K63/02—Receptacles specially adapted for transporting live fish
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
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- A61K31/215—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids
- A61K31/235—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group
- A61K31/24—Esters, e.g. nitroglycerine, selenocyanates of carboxylic acids having an aromatic ring attached to a carboxyl group having an amino or nitro group
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- A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
- A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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Abstract
The invention discloses a method for compound keeping-alive of a slow-release anesthetic, which comprises the steps of temporarily culturing live fish for 1-7 days before transportation; putting the fish into a program temperature control and living device, adding 30-50mg/L slow-release anesthetic into a water body, and adjusting the water temperature from 22-28 ℃ to the subcritical storage temperature of 1-4 ℃ according to the cooling rate of 1-5 ℃/h before transportation; and continuously keeping the temperature of 1-4 ℃ in the transportation process; and (3) after the fish arrives at the destination, putting the fish into 1-4 ℃ culture water, heating the culture water to 22-28 ℃ according to the heating rate of 1-5 ℃/h, and finishing the keep-alive. The survival method of the invention combines the critical dormancy temperature with the slow-release anesthetic, which not only can effectively solve the high stress reaction and high death rate of the fish in the transportation process, but also can improve the stability of the anesthesia effect and the persistence of the anesthesia time, simultaneously can effectively reduce the serum lactic acid concentration and the blood sugar concentration in the fish meat, and obviously improves the cooking loss rate, the muscle fiber diameter, the color difference and other qualities of the fish meat.
Description
Technical Field
The invention relates to the field of aquatic product transportation, in particular to a method for compound keep-alive of a slow-release anesthetic.
Background
Crucian carp is one of the most common freshwater fishes in China, and is mainly a omnivorous fish taking plants as food, prefers to be gathered and grown, and prefers to be eaten and grown. The meat quality is tender, the nutritive value is high, each hundred grams of meat contains 13 grams of protein and 11 grams of fat, and contains a large amount of minerals such as calcium, phosphorus, iron and the like. The crucian has high medicinal value, is neutral in nature and sweet in taste, enters the stomach and the kidney, and has the effects of regulating the middle warmer, tonifying deficiency, removing food, warming the stomach, eating, tonifying the middle warmer and generating qi. Therefore, the crucian carp is deeply favored by consumers and has higher market value and consumption demand.
China is a large country for fish production and consumption, but the regional distribution of the fish farming industry is unbalanced, so that the fish resources in China need to meet the market demand through long-distance transportation. In the transportation process of the fishes, dealers mostly adopt a high-density loading mode so as to achieve the effects of reducing the transportation cost and improving the transportation volume. High-density transportation of live fish, if the transportation conditions are not properly controlled, causes nearly 10% of live fish to die after arriving at the destination, and seriously affects fish trade and cost control. In addition, the transport and storage conditions of the fish also have an important influence on the eating quality of the fish, and the eating quality of the live fish is generally superior to that of dead fish. Therefore, the research and development of technologies for keeping alive and reducing the death rate in the fish transportation process become the key for promoting the fish trade development. The most used keep-alive methods at present are a low-temperature keep-alive method and an anesthesia keep-alive method, but the low temperature has a slow and unstable stabilization effect on crucian, and the anesthesia keep-alive method using a common anesthetic has the problems of unstable anesthesia effect and short duration or too long drug withdrawal period and residue. Therefore, the compound keep-alive method of the slow-release anesthetic is used for keep-alive transportation.
Based on the analysis, in order to solve the problems that the low temperature has slow and unstable stabilizing effect on the crucian, the general anesthetic used in the anesthesia keep-alive method has unstable anesthetic effect and short duration time or has long drug withdrawal period and residue. Therefore, the compound keep-alive method using the slow-release anesthetic can effectively solve the problems of high stress reaction and high death rate of the fishes in the transportation process, improve the stability of the anesthetic effect and the persistence of the anesthetic time, effectively reduce the concentration of serum lactic acid and the concentration of blood sugar in the fish meat, and obviously improve the quality of the fish meat such as cooking loss rate, muscle fiber diameter, color difference and the like.
Disclosure of Invention
In view of the defects, the invention aims to solve the problems that the low temperature has slow and unstable calming effect on crucian carps, the general anesthetic used in the anesthesia keep-alive method has unstable anesthetic effect and short duration time or has long withdrawal period and residue, and develops a method for compound keep-alive of slow-release anesthetic.
The invention is realized by the following technical scheme:
a method for compound keeping-alive of slow-release anesthetic comprises the following steps:
(1) temporary culture: temporarily culturing the fishes in special temporary culture water 1-7 days before transportation;
(2) preparation of the slow-release anesthetic: dissolving and mixing phospholipid, cholesterol, resistant cyclized starch and MS-222 anesthetic in a solvent system, fully mixing uniformly, freezing, and freeze-drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound into a water body, and putting the fish into a program temperature-control survival device; (ii) a
(4) Controlling temperature by program: before transportation, the water temperature is gradually reduced to the subcritical storage temperature, and the subcritical storage temperature is transported at constant temperature;
(5) and (4) awakening: and after the fish reaches the destination, putting the fish into culture water, heating to 22-28 ℃, and finishing keeping alive.
Further, the special temporary rearing water in the step (1) is prepared by the following method:
filtering and continuously aerating tap water to obtain standby water;
adding vitamin C with concentration of 20-30mg/L into the prepared water to obtain the special temporary culture water.
Further, the filtration is carried out by adopting granular activated carbon; the continuous aeration time is 24-48 h.
Furthermore, the water temperature of the temporary culture water is 22-28 ℃, the dissolved oxygen amount is 4-6mg/L, and the pH value is 7.2-7.8.
Further, the pH value is measured every 3-6 h.
Further, in the method for preparing an extended release anesthetic described in step (2), the phospholipid used is selected from any one or more of myristoyl-sn-glycero-3-phospho- (1' -rac-glycerol) (sodium salt) (DMPG), L- α -phosphatidylcholine (EPC), 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and the solvent system used is selected from any one of tert-butanol and a tert-butanol-water co-solvent. The phospholipid and MS-222 anesthetic are used in a ratio of at least 1:0.5 to 1: 1; the ratio of phospholipid to cholesterol is 1:0.01 to 1: 1; the ratio of phospholipids to resistant cyclized starch used is from 1:0.01 to 1: 1.
Further, the type of the anesthetic in the step (3) is a slow-release anesthetic, and the concentration of the anesthetic is 30-50 mg/L.
Further, the water temperature before transportation in the step (4) is 22-28 ℃, the cooling rate is 1-5 ℃/h, and the subcritical storage temperature is 1-4 ℃.
Further, the water temperature of the aquaculture water in the step (5) is 1-4 ℃, and the heating rate is 1-5 ℃/h.
The invention also discloses fish obtained by the method for compound keep-alive of any slow-release anesthetic.
The invention has the beneficial effects that:
in order to solve the problems that the low temperature has slow and unstable calming effect on crucian carps, the general anesthetic used in the anesthesia keep-alive method has unstable anesthesia effect and short duration or has long withdrawal period and residue. According to the invention, before fish transportation, the fish is temporarily fed with special temporary culture water, meanwhile, an anesthetic is embedded, then a slow-release anesthetic of 30-50mg/L is added into the water, and the compound keep-alive treatment is carried out at the critical dormancy temperature of 1-4 ℃, so that the problems of high stress reaction and high death rate of the fish in the transportation process can be effectively solved, the stability of the anesthetic effect and the continuity of the anesthetic time can be improved, the serum lactic acid concentration and the blood sugar concentration in the fish can be effectively reduced, and the qualities of the fish such as cooking loss rate, muscle fiber diameter, color difference and the like can be obviously improved.
Drawings
Fig. 1 shows the breathing rate of crucian at different temperatures and different time periods.
Detailed Description
In order to make the operational procedures and the creative features of the implementation of the invention easy to understand, the invention is further explained below by combining the specific embodiments.
Example 1
A composite keep-alive method of slow-release anesthetic comprises the following specific operation steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 5 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is firstly 1 ℃/h and then 5 ℃/h; raising the temperature of the culture water to 25 ℃.
Example 2
A composite keep-alive method of slow-release anesthetic comprises the following specific operation steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 3 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 3 ℃/h later; raising the temperature of the culture water to 25 ℃.
Example 3
A composite keep-alive method of slow-release anesthetic comprises the following specific operation steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Example 4
A composite keep-alive method of slow-release anesthetic comprises the following specific operation steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: 1, 2-myristoyl-sn-glycero-3-phosphate- (1' -rac-glycerol) (DMPG), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed as follows: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Example 5
A composite keep-alive method of slow-release anesthetic comprises the following specific operation steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), cholesterol, resistant cyclized starch and MS-222 anesthetic were mixed in 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Comparative example 1
Compared with the embodiment 3, the temporary culture water is not professionally treated, and the rest of the operation is the same as the embodiment 3, and the specific steps are as follows:
(1) temporary culture: temporary culture for 2d before transportation, temporary culture water: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the water temperature of the temporary culture water is 25 ℃, and the dissolved oxygen is 6 mg/L;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Comparative example 2
Compared with the example 3, the MS-222 anesthetic is not subjected to embedding slow release treatment, and the rest of the operation is the same as the example 3, which comprises the following steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 6mg/L, and the pH value is 7.5;
(2) anesthesia: adding 40mg/L MS-222 anesthetic into the water body, and putting the fish into a program temperature control and survival device;
(3) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(4) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(5) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Comparative example 3
Compared with the example 3, the anesthesia treatment is not carried out, and the rest of the operation is the same as the example 3, which comprises the following steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 6mg/L, and the pH value is 7.5;
(2) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the program temperature-control live-keeping device for the live fish is adjusted from 25 ℃ to the subcritical storage temperature of 1.6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(3) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 1.6 ℃ in the transportation process;
(4) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 1.6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Comparative example 4
Compared with the embodiment 3, the keep-alive transportation is carried out outside the critical dormancy temperature range, and the rest of the operation is the same as the embodiment 3, which comprises the following steps:
(1) temporary culture: temporarily culturing for 2 days before transportation, adding vitamin C into the temporary culture pond, wherein the concentration is 25mg/L, the water temperature is 25 ℃, and the water for temporary culture: continuously aerating tap water filtered by granular activated carbon for 24 hours for experiment; the temperature of the temporary culture water is 25 ℃, the dissolved oxygen is 5mg/L, and the pH value is 7.5;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) temperature regulation of the program temperature control keep-alive device: before transportation, the water temperature of the live fish program temperature control keep-alive device is adjusted from 25 ℃ to 6 ℃ according to the cooling rate of firstly 2 ℃/h and then 1 ℃/h;
(5) and (3) transportation: placing the program temperature control keep-alive device on a cold chain transport vehicle for transportation, wherein the program temperature control device keeps the required temperature of 6 ℃ in the transportation process;
(6) and (4) awakening: taking out the fish after the fish arrives at the destination, and putting the fish into culture water with the water temperature of 6 ℃; the temperature rise rate of the culture water is 1 ℃/h firstly and then 2 ℃/h later; raising the temperature of the culture water to 25 ℃.
Comparative example 5
Compared with the embodiment 3, the program temperature control, keep-alive and temperature regulation treatment is not carried out on the fishes, and the rest of the operation is the same as the embodiment 3, which specifically comprises the following steps:
the subcritical keep-alive storage temperature has great influence on keep-alive. When the keep-alive transportation temperature of the crucian is maintained at the subcritical storage temperature, the keep-alive time can be prolonged to the maximum extent, the stress response is reduced, and the fish meat eating quality is improved. To determine the optimum temperature of the crucian during transportation, we monitored the breathing rate of the crucian at 1-25 ℃. As can be seen from fig. 1, when the temperature is in the range of 1-2 ℃, the crucian is in a calm state and the respiratory frequency is below 20 times/min. The lowest breathing rate of crucian when the temperature reached 1.6 ℃, i.e., the subcritical storage temperature was 1.6 ℃, so the transport temperature set in this comparative example was 1.6 ℃.
(1) Determination of subcritical storage temperature: cooling the fish placed in the program temperature control keep-alive device at a cooling rate of 5 ℃/h and then 1 ℃/h, and determining the subcritical storage temperature of the fish to be 1.6 ℃ according to the respiratory state and the physiological state of the fish;
(2) preparation of the slow-release anesthetic: l- α -phosphatidylcholine (EPC), cholesterol, resistant cyclized starch, and MS-222 anesthetic were mixed at a ratio of 2: 1: 1: 2, dissolving and mixing the mixture in a solvent system in a mass ratio, fully and uniformly mixing the mixture, freezing the mixture, and then carrying out freeze drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound 40mg/L into a water body, and putting the fish into a program temperature-control and living-keeping device;
(4) and (3) transportation: transporting at normal temperature of 22-28 deg.C;
(5) and (4) awakening: and taking out the fish after the fish arrives at the destination, and putting the fish into the aquaculture water.
Test example 1
The pH values of the temporary culture water of each group of examples and comparative examples were monitored
The concentration of vitamin C determines the pH value of the temporary water, and the pH value is too high or too low, so that the fish body is not adaptive and the stress response is strong. Comparative example 1, which had a lower pH than the other groups due to the absence of vitamin C, further demonstrated that fish survival was not facilitated at pH < 7. Therefore, the concentration of the vitamin C in the temporary culture water is adjusted to be 20-30mg/L according to the pH range of 7.2-7.8.
Test example 2
Monitoring blood index and tissue index of crucian in each group of examples and comparative examples
In the process of compound keep-alive transportation, the concentration of lactic acid and blood sugar in the crucian serum can reflect the stress reaction degree of the crucian. The content of blood sugar can be increased in the whole keep-alive process, and as can be seen from table 1, the blood sugar value is reduced after the keep-alive process disclosed by the invention is used, so that the transport stress of crucian carp is reduced. Lactic acid is a product of glucose glycolysis under anaerobic conditions to provide energy, most of the lactic acid produced by muscle enters blood, and the increase of the content of lactic acid affects the oxygen transport capacity of hemoglobin in blood, so that the maximum oxygen binding capacity of blood is reduced. The content of lactic acid is obviously reduced after the keep-alive process is used, and the keep-alive transportation is facilitated. The water holding capacity refers to the capacity of keeping original water when the muscle is acted by external force, and is influenced by the deformation and degradation of the myofibrillar protein structure, the tenderness, juiciness, nutrient content, taste and the like of the meat can be influenced by the boiling loss rate, and the water holding capacity is a simple and convenient index for measuring the change condition of the fish meat quality. The boiling loss rate is improved after the keep-alive process is used.
TABLE 1 comparison of blood index and tissue index of each group of crucian carp
Test example 3
Monitoring fish color difference of crucian in each group of examples and comparative example
The meat color is a decisive factor influencing the purchasing behavior of consumers, is an important influencing factor of the shelf life of fresh meat, and can be directly used as an important index for sensory judgment of meat quality. During the storage process, the color of the fish meat is changed due to the occurrence of fat oxidation, protein denaturation, pigment degradation reaction and the like. As can be seen from table 2, the fish meat a and b values of comparative example 2 and comparative example 3, which were not subjected to the embedding anesthetic treatment, were significantly decreased compared to the examples, the fish meat a and b values of comparative example 4, which was subjected to the live-keeping transportation outside the critical resting temperature range, were significantly decreased compared to the examples, and the fish meat L and W values of comparative example 5, which was not subjected to the gradient cooling treatment, were significantly increased compared to the other groups. The fish color of the crucian carp is obviously improved after the keep-alive process is used.
TABLE 2 color difference comparison of crucian flesh
Test example 4
Monitoring of muscle fiber diameter of groups of examples and comparative crucian
The histological characteristics of muscle are the histological basis influencing the quality of muscle, and are one of the important indexes for evaluating the meat quality. Muscle fibers are the basic building blocks of skeletal muscle, and their properties determine the quality of the muscle to some extent. Muscle fiber diameter is an important parameter in characterizing muscle. The diameter of muscle fiber is mainly related to age, nutrition status, variety, weight gain rate and exercise amount. The smaller the diameter of muscle fiber is, the more tender the meat quality is; the larger the muscle fiber diameter, the harder the meat quality. As can be seen from table 3, the test method can keep the diameter of the muscle fiber of the crucian carp meat at a short level and can keep a good loss rate of the fish meat in cooking.
Table 3 comparison of muscle fiber diameters of various groups of crucian carps
According to the indexes, the blood indexes and the tissue indexes of the crucian after being transported for five days show that the cooling rate and the heating rate have great influence on the keep-alive, and whether the MS-222 anesthetic is embedded or not has obvious influence on the keep-alive. Therefore, in the keep-alive process, attention should be paid to effective control of the cooling rate. The temperature reduction rate and the temperature rise rate have great influence on the keep-alive. The slower the cooling rate and the heating rate, the better the keep-alive effect of the fish body. However, due to the excessively slow cooling rate and heating rate, the keep-alive transportation time is too long, so that the investment time in the production process of enterprises is too long, and therefore, it is very important to reasonably select the appropriate cooling rate and heating rate. The temperature reduction rate is 1-5 ℃/h, the temperature rise rate is 1-5 ℃/h, namely, the rate of 2-5 ℃/h is adopted in the temperature range of 25-6 ℃, and the better keep-alive transportation effect can be obtained when the rate of l ℃/h is adopted in the temperature range of 6-1.6 ℃. Whether embedding of MS-222 anesthetic greatly affects survival, the phospholipids in the embedding medium are selected from one or more of 1, 2-myristoyl-sn-glycero-3-phospho- (1' -rac-glycerol) (sodium salt) (DMPG), L- α -phosphatidylcholine (EPC), 1, 2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), and the ratio of phospholipids, cholesterol, resistant cyclized starch, and MS-222 anesthetic is 2: 1: 1: 2, the crucian can enter a deep sedated state with long time and keep normal physiological activities, thereby being beneficial to keep-alive transportation.
After the crucian carp is treated by the keep-alive method, the blood sugar and lactic acid concentration of the crucian carp can be effectively reduced, the stress reaction of the crucian carp is reduced, the normal physiological state of the crucian carp is maintained, and the cooking loss rate, the color difference and the muscle fiber diameter of the crucian carp meat can be improved to a certain extent. The invention has high survival rate after the live-keeping transportation and can greatly reduce the transportation cost. Therefore, the method has important significance for keep-alive transportation.
The above embodiments are illustrative of the principles of the present invention and its efficacy, and are not to be construed as limiting the invention. Any person skilled in the art can modify or change the above-mentioned embodiments without departing from the spirit and scope of the present invention. Accordingly, it is intended that all equivalent modifications or changes which can be made by those skilled in the art without departing from the spirit and technical spirit of the present invention be covered by the claims of the present invention.
Claims (10)
1. A method for compound keeping-alive of slow-release anesthetic comprises the following steps:
(1) temporary culture: temporarily culturing the fishes in special temporary culture water 1-7 days before transportation;
(2) preparation of the slow-release anesthetic: dissolving and mixing phospholipid, cholesterol, resistant cyclized starch and MS-222 anesthetic in a solvent system, fully mixing uniformly, freezing, and freeze-drying to obtain a slow-release anesthetic compound;
(3) anesthesia: adding a slow-release anesthetic compound into a water body, and putting the fish into a program temperature-control survival device;
(4) controlling temperature by program: before transportation, the water temperature is gradually reduced to the subcritical storage temperature, and the subcritical storage temperature is transported at constant temperature;
(5) and (4) awakening: and after the fish reaches the destination, putting the fish into culture water, heating to 22-28 ℃, and finishing keeping alive.
2. A method of extended release anesthetic compound keep alive according to claim 1, wherein:
the special temporary culture water in the step (1) is prepared by the following method:
filtering and continuously aerating tap water to obtain standby water;
adding vitamin C with concentration of 20-30mg/L into the prepared water to obtain the special temporary culture water.
3. A method of extended release anesthetic compound keep alive according to claim 2, wherein:
the filtration is carried out by adopting granular activated carbon;
the continuous aeration time is 24-48 h.
4. A method of extended release anesthetic compound keep alive according to claim 1 or 2, wherein:
the water temperature of the temporary culture water is 22-28 ℃, the dissolved oxygen amount is 4-6mg/L, and the pH value is 7.2-7.8.
5. A composite keep-alive method for slow-release anesthetics as recited in claim 4, wherein:
the pH value is measured every 3-6 h.
6. A method of extended release anesthetic compound keep alive according to claim 1, wherein:
in the preparation of said extended release anesthetic of step (2), the phospholipid used is selected from one or more of 1, 2-myristoyl-sn-glycerol-3-phospho- (1' -rac-glycerol) (sodium salt) (DMPG), L- α -phosphatidylcholine (EPC), 1, 2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), and the solvent system used is selected from one of t-butanol and a t-butanol-water co-solvent.
7. A method of extended release anesthetic compound keep alive according to claim 1, wherein:
in the preparation of the slow-release anesthetic in the step (2), the mass ratio of the phospholipid to the MS-222 anesthetic is 1: 0.5-1: 1; the mass ratio of the phospholipid to the cholesterol is 1: 0.01-1: 1; the mass ratio of the phospholipid to the resistant cyclized starch is 1: 0.01-1: 1.
8. A method of extended release anesthetic compound keep alive according to claim 1, wherein:
the type of the anesthetic in the step (3) is slow-release anesthetic, and the concentration of the anesthetic is 30-50 mg/L.
9. A compound keep-alive method with slow-release anesthetic agent according to claim 1, wherein:
the water temperature before transportation in the step (3) is 22-28 ℃, the cooling rate is 1-5 ℃/h, and the subcritical storage temperature is 1-4 ℃; the water temperature of the aquaculture water in the step (4) is 1-4 ℃, and the heating rate is 1-5 ℃/h.
10. A fish obtained by a method of combined survival with a slow release anaesthetic according to any one of the claims 1-9.
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