Application of combination of corona dormancy and taurine in keep-alive transportation of trachinotus ovatus

Abstract

The invention discloses application of combination of corona dormancy and taurine in keep-alive transportation of trachinotus ovatus. Compared with an existing trachinotus ovatus transportation technology, the trachinotus ovatus is dormant through corona, and the trachinotus ovatus is transported in a keep-alive mode through the protection liquid which is sufficient in oxygen and added with taurine, so that the keep-alive transportation stress reaction of the trachinotus ovatus is effectively relieved, the survival time of the trachinotus ovatus is prolonged by nearly two times, the survival rate of the trachinotus ovatus in the transportation process is further increased, and the trachinotus ovatus transportation efficiency is improved. The nutritional quality of the trachinotus ovatus meat is maintained, the transportation cost is remarkably reduced, the economic benefit of the trachinotus ovatus breeding industry is improved, and the method has important application and popularization value.

Description

Technical Field

[0001] The invention relates to the field of aquatic product live transport, and in particular to application of corona dormancy combined with taurine in the live transport of golden pomfret. Background Art

[0002] Golden pompano is a Perciformes fish, with the scientific name Oval pompano, also known as yellow fish. The fish body is flattened and oval, and the body length is about twice the body height. Golden pompano is a warm-water pelagic fish with a very rich protein content of about 15.6%. Golden pompano has no intramuscular spines, tender meat, delicious taste and is rich in protein, linoleic acid, DHA and other unsaturated fatty acids, which can prevent and treat various cardiovascular and cerebrovascular diseases to a certain extent. Golden pompano is mainly distributed in tropical and subtropical waters such as the East China Sea, the South China Sea, the Yellow Sea and the Bohai Sea in my country. It is the most modernized and intensive marine aquaculture fish in my country, and is also the preferred variety for expanding deep-sea aquaculture space, with huge development potential.

[0003] Golden pomfret is a warm-loving, omnivorous, migratory fish that swims day and night, consumes a lot of oxygen, and has a vigorous metabolism. However, it is not very resistant to low temperatures and has a weak ability to resist stress. High-density concentrated fishing and transportation can easily cause golden pomfret to have a strong stress response, which can easily lead to death in a low-temperature environment (<13°C), making it very difficult to transport it alive. In order to reduce the stress response and mechanical damage of golden pomfret during the process of keeping it alive and transporting it, reduce the metabolic rate, and increase the survival rate during transportation, golden pomfret will be induced to hibernate before transportation, and anti-stress agents will be administered to golden pomfret during transportation.

[0004] At present, golden pomfret is mainly sold in the form of frozen fresh products. Live golden pomfret is hard to find in seafood markets far away from the breeding grounds. Even if it is available, the price is very expensive, which limits the expansion of the sales market. Developing a high-density, low-stress and high-survival rate live transportation method is of great significance to the development of golden pomfret aquaculture and the improvement of economic benefits. Summary of the invention

[0005] In order to solve the above problems existing in the prior art, the present invention provides an application of corona dormancy combined with taurine in the alive transportation of golden pomfret.

[0006] The first object of the present invention is to provide an application of corona dormancy combined with taurine in the alive transportation of golden pomfret.

[0007] The second object of the present invention is to provide an application of corona dormancy combined with taurine in alleviating transportation stress of golden pomfret.

[0008] The third object of the present invention is to provide a method for transporting golden pomfret alive.

[0009] In order to achieve the above object, the present invention is implemented by the following scheme:

[0010] Corona hibernation is a new type of fish hibernation technology established by the inventor in the early stage (DOI: 10.19663 / j.issn2095-9869.20240305002). The present invention is the first to use corona hibernation combined with taurine to keep golden pomfret alive during transportation, which can effectively alleviate transportation stress, prolong the survival time, improve the survival rate, and maintain the appearance and meat quality of golden pomfret.

[0011] Therefore, the present invention requests protection for the following contents:

[0012] The invention relates to an application of corona dormancy combined with taurine in the alive transportation of golden pomfret. The taurine is used to prepare an alive liquid, and the golden pomfret after corona dormancy is placed in the protective liquid for alive transportation.

[0013] Preferably, the live transport is water-based live transport.

[0014] Application of corona hibernation combined with taurine in alleviating transportation stress in golden pomfret.

[0015] Preferably, the method of transportation is keep-alive transportation.

[0016] More preferably, the live transport is water-based live transport.

[0017] A method for keeping golden pomfret alive and transporting the golden pomfret in corona dormancy is disclosed. The method comprises transporting the golden pomfret alive and corona dormant in a protective liquid. The protective liquid is fish farming water containing taurine, wherein the working concentration of the taurine is 20-120 mg / L. The temperature of the fish farming water is 15-25°C.

[0018] Preferably, the working concentration of taurine is 70 mg / L.

[0019] Preferably, the volume mass ratio of the protective liquid to the golden pomfret is (1-3) L:1 kg.

[0020] More preferably, the volume mass ratio of the protective liquid to the golden pomfret is 2L:1kg.

[0021] Preferably, the temperature of the water used for fish farming is 20°C.

[0022] Preferably, the salinity of the water used for fish farming is 20-25‰, and the dissolved oxygen content is 6.0-8.0 mg / L.

[0023] More preferably, the dissolved oxygen content of the water used for fish farming is 6.5-7.5 mg / L.

[0024] Preferably, the fish farming water comprises seawater.

[0025] More preferably, the seawater is fresh seawater.

[0026] Further preferably, the water for fish farming is fresh seawater with a temperature of 15-25°C, a salinity of 20-25‰, and a dissolved oxygen content of 6.0-8.0 mg / L.

[0027] More preferably, the water used for fish farming is fresh seawater with a temperature of 20°C, a salinity of 20-25‰, and a dissolved oxygen content of 6.5-7.5 mg / L.

[0028] Preferably, the electric shock conditions for corona dormancy include: electric shock voltage 130-150V, electric shock temperature 15-25°C, and electric shock time 3-5s.

[0029] More preferably, the shocking conditions for corona dormancy include: shocking voltage 140V, shocking temperature 20°C, and shocking time 4s.

[0030] More preferably, the electric shock conditions for corona dormancy also include: electric shock current 0.5-1.5A, output frequency 25-35Hz, duty cycle 20-30%.

[0031] Further preferably, the electric shock conditions for corona dormancy also include: electric shock current 1A, output frequency 30Hz, duty cycle 25%.

[0032] More preferably, the electric shock conditions for corona dormancy include: electric shock voltage 140V, electric shock temperature 20°C, electric shock time 4s, electric shock current 1A, output frequency 30Hz, and duty cycle 25%.

[0033] Preferably, the golden pomfret is firstly stopped from feeding and temporarily kept for 4 to 8 hours, and then put into corona hibernation.

[0034] More preferably, the golden pomfret is temporarily kept in seawater at 20-25° C. without feeding for 4-8 hours.

[0035] More preferably, the golden pomfret is kept in seawater at 20-25° C. without feeding for 6 hours.

[0036] More preferably, the golden pomfret is kept in seawater at 25° C. without feeding for 4 to 8 hours.

[0037] More preferably, the golden pomfret is kept in seawater at 25° C. without feeding for 6 hours.

[0038] More preferably, the seawater is naturally purified seawater.

[0039] More preferably, the salinity of the seawater is 20-25‰, and the dissolved oxygen content is 6.0-8.0 mg / L.

[0040] Still more preferably, the salinity of the seawater is 20-25‰, and the dissolved oxygen content is 6.5-7.5 mg / L.

[0041] Preferably, the time of the live transport is 0h to 72h.

[0042] More preferably, the time of the live transport is 0h to 48h.

[0043] More preferably, the time of the live-preserving transportation is 24h to 48h.

[0044] More preferably, the time of the live transport is 24h to 36h.

[0045] Still more preferably, the time of the live-keeping transportation is 24 hours.

[0046] Compared with the prior art, the present invention has the following beneficial effects:

[0047] Compared with the existing golden pomfret transportation technology, the present invention stuns dormant golden pomfret and uses a protective liquid with sufficient oxygen and added taurine to keep the golden pomfret alive for transportation, effectively alleviating the golden pomfret's stress response during transportation, extending the survival time of the golden pomfret by nearly two times, and also improving the survival rate of the golden pomfret during transportation, while maintaining the nutritional quality of the fish meat, significantly reducing transportation costs, and helping to improve the economic benefits of the golden pomfret farming industry, and has important promotion and application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0048] Figure 1 The effects of corona hibernation and taurine on serum indicators of golden pomfret; A is glucose content, B is cortisol content, C is aspartate aminotransferase content, and D is reactive oxygen content; different letters marked on the rectangular bars indicate statistical significance after variance analysis, P<0.05.

[0049] Figure 2 The effect of corona hibernation and taurine on the malondialdehyde content in the liver and brain tissues of golden pomfret; different letters marked on the rectangular bars indicate statistical significance after variance analysis, P<0.05.

[0050] Figure 3 The effects of corona hibernation and taurine on superoxide dismutase activity in the liver and brain tissues of golden pomfret; different letters on the rectangular bars indicate statistical significance after analysis of variance, P<0.05.

[0051] Figure 4 The effects of corona hibernation and taurine on the activity of glutathione S-transferase in the liver and brain tissues of golden pomfret; different letters marked on the rectangular bars indicate statistical significance after analysis of variance, P<0.05.

[0052] Figure 5 The effects of corona hibernation and taurine on the muscle glycogen (A) and lactic acid (B) contents of golden pomfret after live transport; different letters marked on the rectangular bars indicate statistical significance after analysis of variance, P<0.05. DETAILED DESCRIPTION

[0053] The present invention is further described in detail below in conjunction with the accompanying drawings and specific examples of the specification. The examples are only used to explain the present invention and are not used to limit the scope of the present invention. The test methods used in the following examples are conventional methods unless otherwise specified; the materials and reagents used are reagents and materials that can be obtained from commercial channels unless otherwise specified.

[0054] The golden pomfret used in the following examples was purchased from the Zhanjiang Seafood Wholesale Market, and the golden pomfret in the wholesale market mainly came from the Hainan golden pomfret breeding base.

[0055] The reagents used in the following examples include: Astragalus polysaccharide, analytical grade; Vitamin C (L-ascorbic acid), analytical grade, all purchased from Shanghai Yien Chemical Technology Co., Ltd. Taurine (2-aminoethanesulfonic acid), analytical grade; Thyme essential oil, analytical grade; Clove essential oil, analytical grade, all purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0056] The equipment used in the following embodiments includes: the corona dormancy device is a YDMZ-Lab type portable fish corona dormancy device with an output voltage of 50 to 990V, an output frequency of 1 to 120Hz, an output power of 25 to 400W, an adjustable power-on area of ​​1000×400mm, and the Institute of Fishery Machinery and Instruments of the Chinese Academy of Fishery Sciences; the live transport system is equipped with a cyclone protein separator, a biomechanical filter, a refrigerator, a PP temporary holding tank with a volume of 1 cubic meter, and a live tank containing an oxygen pump, a circulating pump, and a filter, for static simulated transportation.

[0057] Example 1 Effects of corona hibernation combined with different anti-stress agents on the survival time of golden pomfret transported alive in water

[0058] 1. Transport of golden pomfret alive in water

[0059] (1) Stop eating and rest

[0060] 102 golden pomfret (average weight 500g) with consistent growth were selected and randomly divided into 17 groups (recorded as groups 1 to 17), with 6 fish in each group. Each group of golden pomfret was placed in a temporary holding tank and temporarily held for 6 hours in 1020L natural purified seawater at 25℃, salinity 25‰, and dissolved oxygen content of 7.0-8.0mg / L without feeding.

[0061] (2) Preparation of protective solution

[0062] Fresh seawater: salinity 20-25‰, dissolved oxygen 6.5-7.5 mg / L.

[0063] Anti-stress agents: Vitamin C (working concentrations of 10, 25 or 40 mg / L), astragalus polysaccharides (working concentrations of 25, 50 or 75 mg / L), taurine (working concentrations of 20, 70 or 120 mg / L), thyme essential oil (working concentrations of 5, 10 or 15 mg / L), clove oil (working concentrations of 5, 10 or 10 mg / L).

[0064] Each anti-stress agent was fully dissolved in fresh seawater at 15°C at a corresponding concentration to obtain 15 kinds of protective solutions containing different anti-stress agents at different concentrations.

[0065] (3) Corona dormancy

[0066] After the temporary rearing, the golden pomfret of groups 2 to 17 were placed in an electric shock box filled with 20°C fresh seawater and were shocked with a corona dormancy device. The electric shock parameters were set as follows: electric shock voltage 140V, electric shock current 1A, output frequency 30Hz, duty cycle 25%, and electric shock cycle 4s. The electric shock treatment method was: the body of the golden pomfret was parallel and centered between the positive and negative electrode plates, and the current was allowed to pass through the fish body with the largest area during the electric shock, and a shock cycle was processed. After the electric shock, the golden pomfret of groups 2 to 17 all entered a dormant state.

[0067] (4) Keep-alive transport

[0068] After the temporary rearing is completed, the golden pomfret of Group 1 is directly transferred to the live-keeping box of the live-keeping transportation system for live-keeping transportation.

[0069] After the electroshock hibernation, the golden pomfret of groups 2 to 17 were taken out of the electroshock box and transferred to the survival box of the survival transport system for survival transport. The survival box of group 2 was filled with 15℃ fresh seawater, and the volume mass ratio of fresh seawater to fish body was 2L:1kg; the survival boxes of groups 3 to 17 were respectively filled with 15 kinds of protective liquids, and the volume mass ratio of protective liquid to fish body was 2L:1kg.

[0070] The timing started from the moment the golden pomfret of each group entered the survival box. During the transportation process, the survival of the golden pomfret was observed once every hour. The absence of response to stimulation was considered death. When all the golden pomfret in the group died, the timing was stopped and the transportation ended. The number of deaths and the timing duration of each group of golden pomfret were recorded, and the survival rate and survival time were calculated. After the transportation was completed, the pH of the transport water of each group was measured.

[0071] 2. The result of water-saving transportation

[0072] Table 1 Effects of corona hibernation combined with different anti-stress agents on the survival time of golden pomfret in water transport

[0073]

[0074]

[0075] Note: In the survival time, the same letters indicate no significant difference among the groups (P≥0.05), and different letters indicate significant difference among the groups (P<0.05).

[0076] As shown in Table 1, compared with Group 1 which was directly transported alive without any treatment, Groups 2-17 had significantly longer survival time and higher survival rate, indicating that corona hibernation is beneficial to improving the survival and transportation effect of golden pomfret; compared with Group 2, Groups 3-5 had longer survival time and higher survival rate, but the pH value of the water was lower, indicating that vitamin C can cooperate with corona hibernation to promote the survival of golden pomfret during survival and transportation, but the acidic water corrodes and damages the fish skin, increases the risk of infection, and seriously affects the appearance and quality of golden pomfret after transportation; compared with Group 2 , Groups 6-7 had longer survival time and higher survival rate, while Group 8 had shorter survival time and lower survival rate at 48h, indicating that medium and low concentrations of Astragalus polysaccharides can cooperate with corona dormancy to promote the survival of golden pomfret during survival transportation. However, because Astragalus polysaccharides have a fine brown-yellow powder texture, adding too much will cause turbidity in the water, leading to breathing difficulties for fish and reducing the efficiency of water oxygenation. In addition, after Astragalus polysaccharides combine with the mucus protein on the fish's epidermis, the operation of the oxygenator and circulating pump will increase the foam in the survival equipment, aggravating hypoxia. Therefore, high concentrations of Astragalus polysaccharides can promote the survival of golden pomfret during survival transportation. Astragalus polysaccharide could not play the beneficial role of corona hibernation; compared with group 2, groups 9-11 had longer survival time and higher survival rate, and the survival rate of group 10 was still as high as 50% at 72h, even better than most groups using other anti-stress agents, indicating that taurine can better cooperate with corona hibernation to significantly promote the survival of golden pomfret during live transportation compared with other anti-stress agents; compared with group 2, groups 12-13 had longer survival time and higher survival rate, while group 8 had shorter survival time and lower survival rate at 36 and 48h, indicating that low and medium concentrations of Thyme essential oil can cooperate with corona dormancy to promote the survival of golden pomfret during live transport, but thyme essential oil is volatile. After being added to the water, the content of thyme essential oil gradually decreases with time. If the concentration added is too high, the fish will enter deep dormancy, respiratory arrest, and death from lack of oxygen. Therefore, high concentration of thyme essential oil cannot play the beneficial role of corona dormancy. Compared with Group 2, the survival rate of Groups 15 to 17 was slightly higher at 36 hours, but the survival time was significantly shorter, indicating that clove essential oil inhibited the advantageous effect of corona dormancy in promoting live transport.

[0077] In summary, after the golden pomfret was temporarily kept for 6 hours without feeding, it was treated for 4 seconds in a 20°C water environment with parameters of electric shock voltage of 140V, electric shock current of 1A, output frequency of 30Hz, and duty cycle of 25%. The longest survival time reached 67.7±10.0h under the condition of containing 2 volumes of taurine protective solution, which was 85.99% higher than the survival time of direct alive transportation without any treatment. Taurine has the best survival effect when the concentration is 70mg / L. Example 2 Effects of corona dormancy and taurine on serum biochemical indicators of golden pomfret

[0078] 1. Transport of golden pomfret alive in water

[0079] (1) Stop eating and rest

[0080] 48 golden pomfret (average weight 500g) with consistent growth were selected and randomly divided into 8 groups (referred to as Groups 1 to 8), with 6 fish in each group. Each group of golden pomfret was placed in a temporary holding tank and temporarily held for 6 hours in 960L natural purified seawater at 25℃, salinity 25‰, and dissolved oxygen content of 7.0-8.0mg / L without feeding.

[0081] (2) Preparation of protective solution

[0082] Taurine was dissolved in 15°C fresh seawater (salinity 20-25‰, dissolved oxygen 6.5-7.5 mg / L) to obtain a protective solution with a taurine working concentration of 70 mg / L.

[0083] (3) Group processing

[0084] The golden pomfret in Group 1 was not transported alive after temporary rearing, but was directly sampled for testing and recorded as the fresh group (FG).

[0085] After the temporary rearing, the golden pomfret in the second group was put into hibernation by electric shock according to the method in Example 1, and after entering the hibernation state, they were not transported alive, but directly sampled for testing, and recorded as the hibernation group (DG).

[0086] After the temporary rearing, the golden pomfret in Group 3 and Group 4 were transferred to the live-keeping tank of the live-keeping transport system. The live-keeping tank was filled with 20℃ fresh seawater. After all the golden pomfret were put in, the temperature was lowered by 2℃ per hour until the water temperature dropped to 15℃, and then the live-keeping transport was carried out. Groups 3 and 4 were the control groups. Among them, Group 3 was sampled and tested at 24h of live-keeping transport, recorded as control group 24 (CG24 h); Group 4 was sampled and tested at 36h of live-keeping transport, recorded as control group 36 (CG36 h).

[0087] After the temporary rearing, the dormant golden pomfret of Groups 5 to 8 were stunned according to the method of Example 1, taken out from the shock box, and transferred to the live-keeping box of the live-keeping transportation system for live-keeping transportation. The live-keeping box was filled with the protective solution prepared in this example, and the volume mass ratio of the protective solution to the fish body was 2L:1kg. Groups 5 to 8 were experimental groups, and samples were collected and tested at 24h, 36h, 48h and 72h of live-keeping transportation, respectively, and were recorded as experimental group 24 (EG24h), experimental group 36 (EG36 h), experimental group 48 (EG48 h) and experimental group 72h (EG72 h).

[0088] 2. Sampling and index testing

[0089] After the corresponding treatment, 5 golden pomfret were randomly caught in each group, and the fish tail venous blood was collected using a disposable medical syringe and quickly injected into a blood collection tube without adding anticoagulant. Each fish was operated only once. After standing for 120 minutes, the centrifuge was precooled to 4°C in advance, centrifuged at 3500r / min for 20 minutes, and the supernatant was collected to obtain serum, which was packaged and stored at 4°C for testing.

[0090] Glucose (GLU) kit (F006-1-1, Nanjing Jiancheng Bioengineering Institute), cortisol (COR) assay kit (ml103529, Shanghai ELISA Biotechnology Co., Ltd.), aspartate aminotransferase (GOT) kit (C010-2-1, Nanjing Jiancheng Bioengineering Institute) and reactive oxygen species (ROS) enzyme-linked immunosorbent assay kit (ml092661, Shanghai ELISA Biotechnology Co., Ltd.) were used to detect the levels of glucose (Glucose), cortisol (COR), aspartate aminotransferase (GOT) and reactive oxygen species (ROS) in the serum of each group of golden pomfret.

[0091] The glucose concentration in fish blood is one of the important indicators to measure the level of fish sugar metabolism and reflect the degree of stress in the fish. Factors such as stress response and environmental stress may lead to increased blood sugar levels in fish.

[0092] Cortisol is an organic compound extracted from the adrenal cortex. It has multiple pharmacological effects such as anti-inflammatory, anti-allergic, and anti-shock. It also plays an important role in the physiological and ecological processes of fish. At the same time, a large amount of cortisol COR will be released when the fish body is subjected to external stress factors. It is an evaluation indicator of stress response.

[0093] GOT is a key enzyme in the liver that connects the metabolism of sugars, lipids, and proteins. Under normal circumstances, the activity of GOT in serum is at a low level and relatively stable. However, when the liver is damaged, a large amount of GOT will be released into the blood, causing a significant increase in enzyme activity.

[0094] ROS is used to maintain cell activity, but it is also the most common harmful product of oxidative stress. It is a partially reduced metabolite of oxygen, has strong oxidizing properties, and is an indispensable product in the metabolic process. When fish are subjected to external environmental stress or disease, a large number of free radicals will quickly be produced in the body to produce excessive ROS, which will cause their cells to undergo apoptosis and necrosis, leading to internal oxidative stress damage, which is one of the reasons for the damage caused by transport stress to fish.

[0095] 3. Test results

[0096] like Figure 1 As shown in A and B in the figure, compared with the fresh group, the blood glucose and cortisol concentrations in the dormant group increased by 84.8% (P < 0.05) and 17.9% (P < 0.05), respectively, indicating that the electrically stimulated dormant golden pomfret produced a greater stress response; compared with the fresh group, the blood glucose and cortisol concentrations in the control 24h group increased by 28.6% and 18.3%, respectively, and the blood glucose concentration in the control 36h group decreased by 49.66%, while the cortisol concentration increased, indicating that the golden pomfret will have a greater stress response in a short period of time when it enters a water-less and gradually cooled survival environment from a temporary storage environment, and the stress response is alleviated as the survival time is prolonged; the experimental 24h group and the experimental 36h group The blood glucose concentrations in the experimental 48h group and the experimental 72h group were significantly lower than those in the fresh group and the dormant group, and showed a significant downward trend with the extension of the survival time. The cortisol concentrations in the experimental 24h group and the experimental 36h group were basically equivalent to those in the fresh group. The increase in cortisol concentration in the experimental 48h group was basically equivalent to that in the dormant group, and the cortisol concentration in the experimental 72h group was significantly higher than that in the dormant group, indicating that the addition of taurine greatly reduced the stress response of golden pomfret caused by electrical stimulation. This may be because taurine promotes the anti-stress ability of golden pomfret and blunts the stress response by regulating the stress axis, thereby reducing the blood glucose concentration caused by stress stimulation. Compared with gradient cooling, corona hibernation and taurine can more effectively promote the golden pomfret to adapt to the impact of the survival environment with less water and low temperature. The blood glucose level in the 72h experimental group dropped to the lowest level while the cortisol concentration rose to the highest level. The reason may be that although the corona hibernation and taurine relieved the stress of the golden pomfret in the early stage of live transport, the long-term fasting caused the energy in the golden pomfret to be exhausted, and with the accumulation of ammonia nitrogen in the water body, the water quality became poor, which in turn caused the stress level of the golden pomfret to increase. This is unavoidable in live transport for too long.

[0097] like Figure 1As shown in C, by comparing the GOT levels of the fresh group and the dormant group, it can be seen that electrical stimulation of the golden pomfret will increase the activity of GOT in the blood. The GOT activity of the control 24h group and the control 36h group showed an upward trend, while the GOT activity of the experimental 24h group and the experimental 36h group showed a downward trend; the GOT activity of the control 24h group was significantly higher than that of the fresh group by 11.4% (P < 0.05), and the GOT activity of the control 36h group further increased to 28.61U / L, while the GOT activity of the experimental 36h group was not significantly different from that of the fresh group. However, with the extension of the survival time, the GOT activity of the experimental 48h group and the experimental 72h group continued to increase, and the GOT activity of the experimental 72h group was basically equivalent to that of the control 36h. Figure 1 As shown in D, the comparison of GOT levels between the fresh group and the dormant group shows that corona dormancy significantly reduces the content of ROS in the blood of fish (P < 0.05). The serum ROS concentrations of the control 24h group and the control 36h group showed an upward trend, and the serum ROS concentrations of the experimental 24h group, the experimental 36h group, the experimental 48 group and the experimental 72h group also showed a slight upward trend. However, the serum ROS concentration of the experimental 36h group was reduced by 11.19% compared with the control 36h group (P < 0.05). It shows that with the extension of the preservation time, the ROS in the serum of golden pomfret continues to increase, and the addition of taurine slows down the accumulation rate of ROS. The above results show that long-term preservation makes the water environment gradually worse, and the stress caused by electric shock, low temperature and environment damages the liver of golden pomfret and aggravates the degree of oxidative stress, but taurine can remove reactive oxygen and other oxidative harmful substances in the body or enhance the antioxidant capacity of the body's antioxidant system, thereby protecting golden pomfret from oxidative damage.

[0098] Example 3 Effects of corona hibernation and taurine on biochemical parameters of liver and brain tissues of golden pomfret

[0099] 1. Index detection

[0100] The golden pomfret in the fresh group, dormant group, control 24 h group, control 36 h group, experimental 24 h group, experimental 36 h group, experimental 48 h group and experimental 72 h group of Example 2 were killed after blood collection, and the brain and liver tissues were separated on crushed ice, the blood was washed in pre-cooled saline and the water was absorbed, and then they were quickly frozen with liquid nitrogen and placed in a -80°C refrigerator for testing.

[0101] Total protein (TP) kit (A045-2-2, Nanjing Jiancheng Bioengineering Institute), malondialdehyde (MDA) kit (A003-1-2, Nanjing Jiancheng Bioengineering Institute), superoxide dismutase (SOD) kit (A001-3-2, Nanjing Jiancheng Bioengineering Institute) and glutathione sulfhydryltransferase (GST-S) kit (A004-1-1, Nanjing Jiancheng Bioengineering Institute) were used to detect the levels of malondialdehyde (MDA), superoxide dismutase (SOD) and glutathione sulfhydryltransferase (GST-S) in the brain and liver of each group.

[0102] Live transport can induce oxidative stress in fish. MDA is a product of intracellular lipid peroxidation, which can reflect the degree of oxidative damage in the body and is closely related to the production of excessive reactive oxygen species. Corona hibernation may cause temporary damage to the liver of fish. When the temperature of the fish environment is constantly decreasing, the antioxidant defense ability of the body will be destroyed when the temperature is too low, which will deepen the degree of lipid peroxidation and significantly increase the MDA content in the serum.

[0103] SOD is the main enzyme that removes ROS. It is widely present in animals and plants and can catalyze the reaction between superoxide anions and hydrogen ions to generate H 2 O 2 and O 2 The changes in SOD activity in fish can accurately reflect the status of free radical scavenging and cell damage in the body. 2 O 2 When there is an excess of ROS, the body needs to synthesize various antioxidants such as SOD to maintain a balance between the production and removal of ROS, thereby preventing metabolic disorders or the outbreak of oxidative stress and reducing the degree of damage to the fish.

[0104] GST-S is an enzyme mainly distributed in the liver. It has the dual functions of removing peroxides and detoxifying. It is one of the most important antioxidant systems in fish. It can inactivate exogenous or endogenous toxic molecules and convert them into water-soluble compounds to achieve the effect of detoxification. It can also reduce the toxic active oxygen induced by environmental stresses such as low temperature, high salt, and heavy metals, thereby reducing the damage to cell structure and function and protecting cells from oxidative damage.

[0105] 2. Test results

[0106] like Figure 2As shown in the results, compared with the fresh group, the MDA content in the liver of the dormant group increased significantly (P < 0.05); the MDA content in the liver of the control 24h group rose to the highest value of 823.8nmol / mg, and the MDA content in the liver of the control 36h group decreased to a level comparable to that of the dormant group; while the MDA content in the liver tissue of the experimental 24h group, the experimental 36h group, the experimental 48h group and the experimental 72h group showed a downward trend as time went on, and reached the lowest level at 72h; during the survival period, the MDA content in the brain tissue of each group was relatively gentle, and the MDA content in the brain tissue of the experimental 24h group, the experimental 36h group and the experimental 48h group was lower than that of the fresh group. This indicates that due to electrical stimulation, lack of water, low temperature and environmental changes, a large amount of reactive oxygen species is produced in the body of golden pomfret, which increases the MDA level in its liver tissue. The addition of taurine can remove excessive reactive oxygen species in the body of golden pomfret, slow down the accumulation rate of ROS in the body to avoid lipid peroxidation, and reduce the generation of MDA.

[0107] like Figure 3 As shown in the figure, compared with the fresh group, the SOD activity in the liver and brain tissues of the dormant group increased significantly (P < 0.05); the SOD activity in the experimental 24h group and the experimental 36h group was significantly higher than that in the control 24h group and the control 36h group of the same period, and the SOD activity in the liver and brain tissues of the experimental group reached the maximum value at 24h and 36h, respectively, which was 19.40% and 7.91% higher than that in the dormant group, but with the extension of the survival time, the SOD activity in each tissue of the experimental group showed a downward trend. This shows that due to electrical stimulation, lack of water, low temperature and environmental changes, a large amount of reactive oxygen is produced in the body of golden pomfret. The addition of taurine increases the SOD activity in the fish, removes reactive oxygen and alleviates oxidative stress damage.

[0108] like Figure 4 As shown, compared with the fresh group, the activity of GST-S in the liver of the dormant group increased significantly by 13.4% (P < 0.05), and the activity of GST-S in the brain and liver of the experimental 24h group and the experimental 36h group was significantly higher than that of the control 24h group and the control 36h group in the same period (P < 0.05), and the activity of GST-S in the liver and brain tissues of the experimental group increased to the maximum value of 4658.98 and 1547.72 nmol / g at 36h and 72h, respectively, which was 23.87% and 73.53% higher than that of the fresh group. This indicates that as the survival time goes by, the water environment gradually deteriorates, and the environmental stress brought by low temperature and ammonia nitrogen to the fish body causes the tissues and organs to activate GST-S to cope with the stress-induced oxidative stress and ammonia nitrogen toxicity, and taurine can promote the activation of GST-S.

[0109] Example 4 Effects of corona hibernation and taurine on biochemical parameters of muscle tissue of golden pomfret

[0110] 1. Index detection

[0111] The golden pomfret in the fresh group, dormant group, control 24 h group, control 36 h group, experimental 24 h group, experimental 36 h group, experimental 48 h group and experimental 72 h group of Example 2 were killed after blood collection, and the back muscle tissue was separated on crushed ice, the blood was washed in pre-cooled saline and the water was absorbed, and then it was quickly frozen with liquid nitrogen and placed in a -80°C refrigerator for testing.

[0112] The basic nutrients represent the inorganic matter, energy components and composition of the muscle and the substances that make up the body and tissue repair. The crude ash content in the muscle of each group of golden pomfret was determined by referring to the first method (burning method) of national standard GB 5009.4-2016; the crude fat content in the muscle of each group of golden pomfret was determined by referring to the Soxhlet extraction method of national standard GB 5009.6-2016; the moisture content in the muscle of each group of golden pomfret was determined by referring to the first method (direct drying method) of national standard GB5009.3-2016; the crude protein content in the muscle of each group of golden pomfret was determined by referring to the Kjeldahl method of national standard GB5009.5-2016.

[0113] The texture characteristics of fish meat reflect its taste quality during consumption, and its changes are affected by the protein content. The hardness, resilience, adhesiveness, elasticity, chewiness and shear force of the muscles of each group of golden pomfret were tested using a texture analyzer (physical property tester TA.XT Plus).

[0114] The changes in glycogen and lactic acid content can reflect the body's use of energy metabolism substances. Glycogen kit (A043-1-1, Nanjing Jiancheng Bioengineering Institute) and lactic acid kit (A019-3-1, Nanjing Jiancheng Bioengineering Institute) were used to detect the content of glycogen and lactic acid in the muscles of each group of golden pomfret.

[0115] 2. Experimental results

[0116] Table 2 Effects of corona hibernation and taurine on basic nutritional components of muscle tissue of golden pomfret

[0117]

[0118] Note: The same letters in the table indicate no significant difference among the groups (P≥0.05), and different letters indicate significant difference among the groups (P<0.05).

[0119] As shown in Table 2, the crude ash, fat, water and protein in the fish meat of each group showed a downward trend during the preservation process. The four nutrients in the control 36h group decreased by 9.94%, 28.08%, 14.85% and 34.76% respectively compared with the fresh group, and were significantly lower than the experimental 24h group, the experimental 36h group and the experimental 48h group (P < 0.05). The four nutrients in the experimental group all dropped to the lowest level at 72h preservation. It may be that the ultra-long fasting preservation transportation still leads to the aggravation of multiple stresses, thus leading to the reduction of nutrients. The above results show that the preservation transportation technology of corona dormancy and taurine can reduce the stress level of golden pomfret and reduce the consumption of nutrients.

[0120] Table 3 Effects of corona dormancy and taurine on the texture characteristics of golden pomfret during live transportation

[0121]

[0122]

[0123] Note: The same letters in the table indicate no significant difference among the groups (P≥0.05), and different letters indicate significant difference among the groups (P<0.05).

[0124] As shown in Table 3, the recovery, adhesiveness, elasticity, chewiness, shear force and other indicators of the experimental group at 24h, 36h and 48h of live transport were not significantly different from those of the fresh group; while the hardness, elasticity, chewiness and other indicators of the control group at 24h and 36h of live transport were significantly reduced compared with the fresh group. This shows that the live transport technology of corona dormancy and taurine can effectively maintain the fish quality of golden pomfret during live transport.

[0125] like Figure 5 As shown in A and B in the figure, with the extension of the preservation time, the glycogen content in the muscles of the control group and the experimental group showed an overall downward trend. The muscle glycogen of the control group and the experimental group dropped to significantly lower levels than the fresh group at 24h and 48h of preservation, respectively (P < 0.05), while the change trend of lactic acid content in the control group and the experimental group was opposite to that of glycogen. This shows that during the long-term fasting preservation process, the glucose in the fish body cannot meet its needs in time, and anaerobic metabolism is carried out, resulting in glycogen decomposition to produce lactic acid, and the preservation transportation technology of corona hibernation and taurine can inhibit glycogen decomposition during the preservation transportation period and retain more glycogen.

[0126] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit the protection scope of the present invention. For ordinary technicians in this field, other different forms of changes or modifications can be made based on the above descriptions and ideas. It is not necessary and impossible to list all the implementation methods here. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the claims of the present invention.

Claims

1. Application of corona dormancy combined with taurine in the alive transportation of golden pomfret.

2. Application of corona hibernation combined with taurine in alleviating transportation stress in golden pomfret.

3. A method for transporting golden pomfret alive, characterized in that: The corona-hibernated golden pomfret is transported alive in a protective solution; the protective solution is fish farming water containing taurine, the working concentration of the taurine is 20-120 mg / L; the temperature of the fish farming water is 15-25°C.

4. The method according to claim 3, characterized in that The volume mass ratio of the protective liquid to the golden pomfret is (1-3) L:1 kg.

5. The method according to claim 3, characterized in that: The salinity of the water for fish farming is 20-25‰, and the dissolved oxygen content is 6.0-8.0 mg / L.

6. The method according to claim 3, characterized in that: The water for fish farming includes sea water.

7. The method according to claim 3, characterized in that The electric shock conditions for the corona dormancy include: an electric shock voltage of 130 to 150 V, an electric shock temperature of 15 to 25° C., and an electric shock cycle of 3 to 5 seconds.

8. The method according to claim 7, characterized in that The electric shock conditions for the corona dormancy also include: electric shock current 0.5-1.5A, output frequency 25-35Hz, duty cycle 20-30%.

9. The method according to claim 3, characterized in that: The time of the live transport is 0h to 72h.

10. The method according to claim 3, characterized in that The golden pomfret is firstly kept without food for 4 to 8 hours and then put into corona hibernation.

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