Breeding method for slowing down escape behavior of jian carps

Abstract

The invention relates to the technical field of aquaculture, in particular to a breeding method for slowing down the escape behavior of jian carps. By adding a certain proportion of tryptophan into the fish feed, the fish feed suitable for rice field culture is provided, the escape behavior of jian carps is effectively slowed down, double harvest of rice and fish is achieved, economic benefits are improved, and a new method is provided for improving the rice field culture technology.

Description

Technical Field

[0001] The invention relates to the technical field of aquaculture, and in particular to a breeding method for slowing down the escape behavior of Jian carp. Background Art

[0002] Farming carp in rice fields is an ecological agricultural model that combines rice cultivation with aquaculture. It can not only improve the utilization rate of land and water resources, but also achieve a bumper harvest of rice and fish, and promote farmers' income. However, the structural defects of traditional breeding facilities are one of the main reasons for carp escapes. For example, the unreasonable mesh specifications of the cages, the insufficient height of the anti-escape wall of the pond, and the poor sealing of the joints provide opportunities for carp to escape. At the same time, changes in the breeding environment, such as deterioration of water quality, stress reactions caused by excessive breeding density, and foraging impulses caused by untimely feeding, will also prompt carp to frequently try to escape. In addition, carp itself has the habits of upstream swimming and jumping. When stimulated by water flow or frightened, the probability of escape increases significantly.

[0003] Although some farmers have adopted conventional measures such as reinforcing fences and increasing the density of cages, these methods have the problems of high cost, difficult maintenance, and great restrictions on the growth environment of carp. Moreover, traditional physical anti-escape methods are difficult to fundamentally solve the escape behavior of carp caused by environmental stress. Therefore, in-depth research on the inducing mechanism of carp escape behavior, combining biological characteristics with engineering technology, and developing a low-cost, high-efficiency comprehensive anti-escape technology system have become key issues that need to be urgently addressed in the current carp farming industry. Summary of the invention

[0004] In view of this, the present invention provides a breeding method for slowing down the escape behavior of Jian carp.

[0005] The invention provides application of tryptophan or fish feed containing tryptophan in slowing down the escape behavior of carp.

[0006] In a specific embodiment of the present invention, an escape test was conducted using Jian carp as an example. The results showed that a specific proportion of tryptophan can significantly reduce the number of fish escapes, and the level of stress-related enzyme activity in serum is reduced, such as malondialdehyde (MDA) and SOD (superoxide dismutase) levels. It shows that adding a specific proportion of tryptophan to fish feed can simply and efficiently slow down the escape behavior of fish in rice field farmed fish, and effectively reduce oxidative stress damage, promote fish growth, and greatly improve fish production and farming economic benefits. In the application of the present invention, the carp includes Jian carp. Furthermore, the Jian carp is Jian carp No. 2.

[0007] In the application of the present invention, the fish feed comprises the following raw materials in percentage by mass:

[0008] Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9% - 30.1%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 0.4% - 1.6%, choline chloride 0.5%, DMPT 0.5%, and soybean oil 8%.

[0009] In some embodiments, the fish feed comprises raw materials in the following mass fractions:

[0010] Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9% - 27.3%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 0.8% - 1.2%, choline chloride 0.5%, DMPT 0.5%, and soybean oil 8%.

[0011] In some specific embodiments, the fish feed comprises raw materials in the following mass fractions:

[0012] Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 27.3%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 1.2%, choline chloride 0.5%, DMPT 0.5%, and soybean oil 8%.

[0013] In other specific embodiments, the fish feed comprises raw materials in the following mass fractions:

[0014] Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 1.6%, choline chloride 0.5%, DMPT 0.5%, and soybean oil 8%.

[0015] Experiments show that in the treatment group fed with the fish feed with the highest addition amount of tryptophan, the activities of SOD and MDA decreased significantly, indicating that there was no increase in enzyme activity caused by oxidative stress. It shows that adding 1.6% tryptophan to the fish feed can effectively reduce oxidative stress damage, thereby efficiently slowing down the escape behavior of fish in paddy field aquaculture. This provides a theoretical basis for the application and promotion of tryptophan in aquatic animal feed and a new way to reduce the escape reaction during the cultivation of Cyprinus carpio var. Jian.

[0016] In the present invention, there are no special restrictions on the specific types of minerals and vitamins, and common commercially available products in the field of aquaculture can be used. The main components of the minerals are trace elements such as calcium bicarbonate, sodium chloride, potassium chloride, and magnesium sulfate. The vitamins include vitamin A, vitamin D, vitamin E, vitamin K3, etc. In the specific embodiments of the present invention, both the minerals and vitamins are premixes and are all purchased from Beijing Dabeinong Technology Group Co., Ltd. in Jiangsu, China.

[0017] Specifically, the composition and content of vitamins in each kilogram of premix are as follows (in international units or milligrams): Vitamin A 900,000 international units; Vitamin D 250,000 international units; Vitamin E 4,500 milligrams; Vitamin K3 220 milligrams; Vitamin B1 320 milligrams; Vitamin B2 1,090 milligrams; Vitamin B5 2,000 milligrams; Vitamin B6 5,000 milligrams; Vitamin B12 116 milligrams; Pantothenate 1,000 milligrams; Folic acid 165 milligrams; Choline 60,000 milligrams; Biotin 50 milligrams; Niacin 2,500 milligrams.

[0018] Specifically, the composition and content of minerals in each kilogram of premix (in grams): Calcium hydrogen phosphate 20 grams; Sodium chloride 2.6 grams; Potassium chloride 5 grams; Magnesium sulfate 2 grams; Ferrous sulfate 0.9 grams; Zinc sulfate 0.06 grams; Copper sulfate 0.02 grams; Manganese sulfate 0.03 grams; Sodium selenite 0.02 grams; Cobalt chloride 0.05 grams; Potassium iodide 0.004 grams.

[0019] The present invention also provides a fish feed for reducing or slowing down the escape of common carp, and the formula composition of the fish feed is as described above.

[0020] The present invention investigated the effect of adding a certain proportion of tryptophan to fish feeds with different formulas on the escape behavior of carp. The results showed that the fish feed provided by the present invention was significantly better than other feeds in reducing the escape of carp. It shows that the formula design of the fish feed of the present invention is scientific and reasonable, can accurately meet the growth habits and behavioral needs of carp in the paddy field culture environment, and can be widely applied to the culture of carp in paddy fields.

[0021] The present invention also provides a paddy field culture method for slowing down the escape behavior of carp, using the fish feed described above as the daily diet to feed carp.

[0022] In some embodiments, the feeding time is 60 days, feeding twice a day, feeding according to the weight of the fish at that time, and the feeding amount each time is 4% - 5% of the weight of the fish. In the specific embodiments of the present invention, the feeding time is 60 days in total, and the method of feeding to satiety is adopted, and the feeding amount is continuously adjusted as the fish weight increases. At the beginning of the experiment, the weight of the experimental fish was 8 - 9 g, and the feeding amount was about 5% of its weight (i.e., 0.45 g / fish), and the experimental fish were grouped and raised, with 10 fish in each bucket and 3 buckets in parallel for each group. After one month of feeding, the weight of the experimental fish was about 25 g, and the feeding amount was about 4% of the weight (i.e., 1 g / fish), and after 60 days of feeding, the weight of the experimental fish was about 45 g, and the feeding amount was approximately 1.8 g / fish.

[0023] Specifically, the two feedings are specifically: feeding once at 8:00 and once at 17:00 respectively.

[0024] The present invention provides a fish feed suitable for paddy field aquaculture by adding a certain proportion of tryptophan to the fish feed, effectively slowing down the escape behavior of Cyprinus carpio var. Jian, achieving a double harvest of rice and fish, improving economic benefits, and providing a new method for improving paddy field aquaculture technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 Shows the test flow chart of the present invention.

[0026] Figure 2 Shows the signal pathways of glycine, serine and threonine metabolism; where 2.1.2.1 corresponds to SHMT, 5.4.2.1.1 corresponds to PGAM, 2.1.4.1 corresponds to GATM, and 1.5.8.4 corresponds to DMGDH.

[0027] Figure 3 Are the qRT-PCR verification results of SHMT, PGAM, GATM, and DMGDH.

[0028] Figure 4 Is the signal pathway of HIF-1; where the English names within the boxes all represent gene names, and signalingpathway is the signal pathway. Among them, GF corresponds to INS, RTK corresponds to INSR, and HIF-1α corresponds to HIF1A.

[0029] Figure 5 Are the qRT-PCR verification results of INSR, HIF1A, ENO1, and INS. DETAILED DESCRIPTION OF THE EMBODIMENTS

[0030] The present invention provides a breeding method for slowing down the escape behavior of Cyprinus carpio var. Jian. Those skilled in the art can draw on the content of this article and appropriately modify the process parameters to achieve it. It should be particularly noted that all similar substitutions and modifications are obvious to those skilled in the art, and they are all considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments, and those skilled in the art can obviously make changes or appropriate modifications and combinations to the methods and applications in this article without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0031] In this article, expressions such as "including", "having", "containing" and their grammatical synonyms should all be understood as open expressions with inclusiveness rather than exclusiveness, and at the same time provide the solution of "consisting of...".

[0032] In addition, the numerical ranges involved in this article, in addition to the disclosed point values, also cover all intermediate values between any two end point values, including any value between any two point values and any small range within any two numerical ranges.

[0033] The test materials used in this invention are all ordinary commercially available products and can be purchased in the market.

[0034] The following further elaborates the present invention in combination with embodiments. The experimental procedure is shown in Figure 1 :

[0035] Example 1

[0036] The experimental fish were taken from Jianli No. 2 of the Freshwater Fisheries Research Center, Chinese Academy of Fishery Sciences, and a 60-day breeding experiment was conducted. The breeding experiment was carried out in 12 breeding barrels (water temperature 20±1°C, pH 7.6±0.2). 120 healthy Jianli fish (10.00±0.5 g) without diseases and injuries were selected and divided into 4 groups (Feed Group 1, Feed Group 2, Feed Group 3, and Feed Group 4), with 3 replicates in each group and 10 fish in each replicate. The fish in Feed Groups 1-4 were fed Feed 1-4 shown in Table 1 in sequence. During the experiment, the 4 feeds were respectively fed to satiety at 8:00 and 17:00 every day.

[0037] The compositions of the 4 feeds are shown in Table 1.

[0038] Table 1

[0039]

[0040] Note: In Table 1, % represents the mass percentage of each raw material in the total raw materials.

[0041] After measurement, the total tryptophan contents in Feeds 1-4 were 0.7%, 1.1%, 1.5%, and 2.1% respectively.

[0042] After the breeding experiment, an 18-day escape experiment was carried out. A special experimental device ("A device and method for measuring fish behavior", Patent No.: ZL2022113332512.2) was used in the escape experiment to simulate the water flow in terraced fields under heavy rain and monitor the escape responses of the experimental fish. Before the experiment, the experimental fish were acclimated in the adaptation tank of the special device for one week. The water temperature of the device was maintained at about 20°C, and oxygen was added to the adaptation tank to stabilize the dissolved oxygen. The formal escape experiment was repeated 3 times. Each replicate was separately placed into the device for the experiment, and the escape rates of the experimental fish in each group were recorded. The results are shown in Table 2. At the same time, the water temperature and feeding conditions were recorded. Under hypoxia and rising water conditions, the escape responses of Jianli fish that would not be carried away by the water flow and had a strong willingness to migrate upstream were weak.

[0043] Table 2 Effects of feeding feeds with different proportions of tryptophan on the escape rate

[0044]

[0045] After the escape experiment, the brains and livers of the experimental fish were taken for subsequent transcriptome and metabolome sequencing,

[0046] The serum was taken for the determination of stress-related indicators, and the results are shown in Table 3.

[0047] Among them, superoxide dismutase (SOD) was determined by the WST-1 method using a superoxide dismutase assay kit purchased from Nanjing Jiancheng Bioengineering Institute; malondialdehyde (MDA) was determined by the TBA method using a malondialdehyde test kit purchased from Nanjing Jiancheng Bioengineering Institute.

[0048] Table 3 Content of related substances

[0049]

[0050] Example 2

[0051] The brains and livers of experimental fish in Feed Group 1 (tryptophan addition amount 0.4%) and Feed Group 4 (tryptophan addition amount 1.6%) were used for transcriptome and metabolome sequencing to analyze differential genes and differential metabolites.

[0052] The results showed that in the biosynthetic signaling pathway of amino acids, the content of tryptophan metabolites in the experimental group (tryptophan addition amount 1.6%) was increased compared with the control group (tryptophan addition amount 0.4%), and the expression of upstream related genes was up-regulated. After verification by qRT-PCR, the results were consistent with the transcriptome results, as shown in Figures 2 - 3 .

[0053] The HIF-1 (hypoxia-inducible factor-1) signaling pathway is a key mechanism for cells to respond to hypoxic stress and also plays an important role under various other stress conditions. HIF-1α is a subunit that composes HIF-1. Under hypoxic conditions, HIF-1α stably translocates to the nucleus, binds to HIF-1β, and activates the transcription of downstream genes.

[0054] According to the sequencing results, the expressions of related genes INSR, HIF1A (i.e., HIF1α), ENO1, and INS in the experimental group (tryptophan addition amount 1.6%) were significantly up-regulated compared with the control group (tryptophan addition amount 0.4%). It was indicated that the ability of the experimental group to respond to stress was enhanced ( Figure 4 ). After verification by qRT-PCR, the results were consistent with the transcriptome results ( Figure 5 ).

[0055] Through metabolome analysis, the differences in liver metabolites of common carp at different tryptophan levels were detected. Combining with transcriptome analysis, differential target genes and their enriched signaling pathways were screened. Using qRT-PCR verification, the molecular mechanism of tryptophan affecting the escape response of common carp was analyzed.

[0056] The results showed that, compared with the other three groups of feeds, feeding Feed 4 could significantly reduce the number of fish escapes, and the levels of enzyme activities related to stress in the serum decreased, such as the levels of malondialdehyde (MDA) and SOD (superoxide dismutase), which was more conducive to the growth of Cyprinus carpio var. Jian.

[0057] The above are only the preferred embodiments of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. Application of tryptophan or fish feed containing tryptophan in slowing down the escape of carp.

2. The application according to claim 1, characterized in that, The carp includes Cyprinus carpio var. Jian.

3. The application according to claim 2, wherein the Cyprinus carpio var. Jian is Cyprinus carpio var. Jian No.

2.

4. The application according to any one of claims 1 to 3, characterized in that The fish feed comprises raw materials in the following mass percentages: Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9% - 30.1%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 0.4% - 1.6%, choline chloride 0.5%, DMPT 0.5% and soybean oil 8%.

5. The application according to claim 4, wherein The fish feed comprises raw materials in the following mass fractions: Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9% - 27.3%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 0.8% - 1.2%, choline chloride 0.5%, DMPT 0.5% and soybean oil 8%.

6. The application according to claim 5, characterized in that The fish feed comprises raw materials in the following mass fractions: Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 27.3%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 1.2%, choline chloride 0.5%, DMPT 0.5% and soybean oil 8%; or Fish meal 5%, casein 28.5%, gelatin 4%, wheat starch 26.9%, microcrystalline cellulose 19.5%, calcium dihydrogen phosphate 2%, vitamins 1%, minerals 0.5%, tryptophan 1.6%, choline chloride 0.5%, DMPT 0.5% and soybean oil 8%.

7. Fish feed for slowing down the escape of Cyprinus carpio var. Jian, characterized in that, It is the fish feed in the application according to any one of claims 4 - 6.

8. A paddy field farming method for slowing down the escape of carp, characterized in that, Feeding carp with the fish feed according to claim 7 as the daily ration.

9. The rice field cultivation method according to claim 8, characterized in that, The feeding time is 60 days, feeding twice a day, and the feeding amount each time is 4% - 5% of the fish body weight.

10. The rice field farming method according to claim 9, characterized in that, Feed once at 8:00 and once at 17:00 respectively.

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