Application of arginine processing waste liquid in aquatic feed additive

Abstract

The invention relates to an application of arginine processing waste liquid in an aquatic feed additive. A powdery compound amino acid chelate prepared by chelation reaction of a pretreated concentrated solution of the arginine processing waste liquid and metal salt is used as the aquatic feed additive; the pretreatment method of the arginine processing waste liquid specifically comprises the following steps: filtering the arginine processing waste liquid through a filter membrane to remove suspended matters, carrying out precipitation reaction by adopting sodium carbonate or sodium bicarbonate, centrifuging to remove solids, and concentrating to obtain a concentrated solution. The compound amino acid chelate prepared by the preparation method disclosed by the invention is added into a basal feed of crabs, has a certain function of promoting growth and weight gain of the crabs, and also can relieve stress response of the crabs under the stress of exposure and improve the survival rate of the crabs under the stress of exposure.

Description

Technical Field

[0001] The present invention belongs to the technical field of feed additives, and specifically relates to the application of arginine processing waste liquid in aquatic feed additives. Background Art

[0002] Arginine processing waste liquid is a by-product generated during the production of arginine through processes such as fermentation or protein hydrolysis extraction. These waste liquids usually contain a certain amount of arginine, other amino acids, organic matter, heavy metal ions, etc. If directly discharged, it will cause waste of resources and environmental pollution.

[0003] Amino acid chelates are compounds formed by the coordination bond combination of amino acids and metal ions, and are widely used in fields such as biochemistry, nutrition, pharmacy, agriculture, and industry; traditional methods (such as CN103553740A) mostly use pure amino acids and metal salts for direct chelation, with high costs and unable to fully utilize industrial waste; based on the starting point of sustainable development and making full use of arginine processing waste liquid, amino acid chelates can be prepared by chelation reaction of arginine processing waste liquid with metal ions. Currently, there is little research on the beneficial effects of amino acid chelates prepared from arginine processing waste liquid as aquatic feed additives on aquatic products (such as crabs). Therefore, the present invention provides an application of arginine processing waste liquid in aquatic feed additives. Summary of the Invention

[0004] The purpose of the present invention is to solve the above problems and provide an application of arginine processing waste liquid in aquatic feed additives.

[0005] The present invention achieves the above purpose through the following technical solutions: The present invention provides an application of arginine processing waste liquid in aquatic feed additives, using the powdery composite amino acid chelate obtained by the chelation reaction of the concentrated solution after pretreatment of arginine processing waste liquid with metal salts as an aquatic feed additive; The pretreatment method of the arginine processing waste liquid is specifically to filter the arginine processing waste liquid through a filter membrane to remove suspended matter, perform a precipitation reaction with sodium carbonate or sodium bicarbonate, centrifuge to remove solids, and concentrate to obtain a concentrated solution.

[0006] As a further optimization scheme of the present invention, the pore size of the filter membrane ≤ 0.45 µm.

[0007] As a further optimization scheme of the present invention, the precipitation reaction is specifically as follows: dissolve sodium carbonate or sodium bicarbonate in deionized water to obtain a sodium carbonate solution or a sodium bicarbonate solution respectively, adjust the pH of the arginine processing waste liquid to 8-11, add the sodium carbonate solution or the sodium bicarbonate solution to the arginine processing waste liquid, continuously stir for 30-40 min and then let it stand to precipitate solids, wherein the addition amount of the sodium carbonate or the sodium bicarbonate in the arginine processing waste liquid is 20-35 g / L.

[0008] As a further optimization scheme of the present invention, the content of the compound amino acids in the concentrated solution is ≥50 g / L.

[0009] As a further optimization scheme of the present invention, the chelation reaction is specifically as follows: adjust the initial pH of the reaction solution of the chelation reaction to 5.5-6.5, under the temperature condition of 70-85 °C, stir at a speed of 800-1500 r / min, and the chelation reaction time is 2-3 h.

[0010] As a further optimization scheme of the present invention, the metal salt is at least one of ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate.

[0011] As a further optimization scheme of the present invention, when the metal salt is a combination of ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate, the molar ratio of Fe 2+ , Cu 2+ , Zn 2+ , Mn 2+ is 1:0.5:2:0.5.

[0012] As a further optimization scheme of the present invention, in the preparation raw materials of the compound amino acid chelate, the molar ratio of the total amount of metal ions in the metal salt to the compound amino acids in the concentrated solution is 1:2-3.

[0013] As a further optimization scheme of the present invention, the dosage of the compound amino acid chelate as an aquatic feed additive is 0.1 wt%-5 wt%.

[0014] As a further optimization scheme of the present invention, the compound amino acid chelate as an aquatic feed additive can promote the growth of crabs, improve the protein utilization rate of crabs, reduce the stress response of crabs under dry exposure stress, and improve the survival rate of crabs under dry exposure stress.

[0015] The beneficial effects of the present invention are as follows: 1) The compound amino acid chelate prepared by the present invention is added to the basic feed of crabs, which has a certain function of promoting the growth and weight gain of crabs, and also has a certain function of improving the protein utilization efficiency of Eriocheir sinensis. 2) The compound amino acid chelate obtained by the preparation of the present invention is added to the basic feed of crabs, and it has a particularly significant effect on promoting the growth and weight gain of Eriocheir hepuensis and improving the protein utilization efficiency of Eriocheir hepuensis. It has a specific effect. 3) The compound amino acid chelate obtained by the preparation of the present invention is added to the basic feed of crabs, which can significantly reduce the serum cortisol content of Eriocheir hepuensis under the stress of dry exposure, indicating that it can relieve the stress response of Eriocheir hepuensis. It can also significantly improve the survival rate of Eriocheir hepuensis under dry exposure stress, has practical application value, can reduce the mortality rate of Eriocheir hepuensis during transportation, and improve economic benefits. During the pretreatment process of the arginine processing waste liquid of the present invention, a precipitation reaction is added, so that the finally prepared compound amino acid chelate can further promote the growth and weight gain of Eriocheir hepuensis and significantly improve the protein utilization efficiency of Eriocheir hepuensis. Specific Embodiments

[0016] The following further describes the present application in detail. It is necessary to point out here that the following specific embodiments are only used to further illustrate the present application and cannot be understood as limiting the protection scope of the present application. Those skilled in the art can make some non-essential improvements and adjustments to the present application based on the above application content.

[0017] I. Materials 1. The composition of the arginine processing waste liquid of the present invention is shown in Table 1: Table 1 Composition Table ; The arginine processing waste liquid of the present invention is the waste liquid generated during the production of arginine by the conventional fermentation method; 2. Hairy crabs: Eriocheir sinensis ( Chinese mitten crab ), Eriocheir hepuensis ( Eriocheir hepuensis ), Eriocheir japonica ( Japanese mitten crab ); 3. Swimming crabs: Portunus trituberculatus ( Swimming crab ), Portunus sanguinolentus ( Portunus sanguinolentus ); 4. The pH regulator is NaOH or HCl; 5. Sodium carbonate (or sodium bicarbonate) solution with a mass concentration of 5%-10%: The sodium carbonate (or sodium bicarbonate) solution obtained by dissolving sodium carbonate (or sodium bicarbonate) in deionized water; the sodium carbonate solution with a mass concentration of 5% is used in the following experiments.

[0018] The methods used in the present invention are all conventional methods known to those skilled in the art unless otherwise specified. The raw materials and reagents used, such as those not otherwise specified, are all commercially available products.

[0019] II. Method 2.1 Preparation of Compound Amino Acid Chelate Example 1 2.1.1 The waste liquid from arginine processing is filtered through a filter membrane with a pore size of 0.45 µm to remove suspended solids, and concentrated to obtain a compound amino acid content of 60 g / L; 2.1.2 Add compound metal salts (ferrous sulfate, copper sulfate, zinc sulfate, manganese sulfate, where the molar ratio of Fe 2+ 、Cu 2+ 、Zn 2+ 、Mn 2+ is 1:0.5:2:0.5) to the concentrated liquid in step 2.1.1, and the molar ratio of the total metal ions to the compound amino acids is 1:2; adjust the pH to 5.8, and under the temperature condition of 80 °C, stir at a speed of 1200 r / min. After chelation reaction for 2.5 h, a reaction solution is obtained; 2.1.3 The reaction solution is centrifuged to remove impurities to obtain a filtrate, and the filtrate is spray-dried (the inlet temperature is 160 - 180 °C, preferably 170 °C, and the outlet temperature is 70 - 90 °C, preferably 80 °C) to make a powdery compound amino acid chelate, with a compound amino acid content of 70.2% and a total metal content of 15.3%.

[0020] Example 2 Based on Example 1, in this example, only the molar ratio of the total metal ions to the compound amino acids in step 2.1.2 is adjusted. In this example, the molar ratio of the total metal ions to the compound amino acids is 1:2.5; the compound amino acid content is 72.3% and the total metal content is 16.8%; the rest are the same as in Example 1.

[0021] Example 3 Based on Example 1, in this example, only the molar ratio of the total metal ions to the compound amino acids in step 2.1.2 is adjusted. In this example, the molar ratio of the total metal ions to the compound amino acids is 1:3; the compound amino acid content is 71.4% and the total metal content is 15.9%; the rest are the same as in Example 1.

[0022] Example 4 Based on Example 2, in this example, only step 2.1.1 is adjusted. In this example, step 2.1.1 is specifically as follows: 1) The waste liquid from arginine processing is filtered through a filter membrane with a pore size of 0.45 µm to remove suspended solids; 2), Sodium carbonate is used for the precipitation reaction. Specifically, the pH of the arginine processing waste liquid is adjusted to 8.5, and a sodium carbonate solution is added to the arginine processing waste liquid (the addition amount of sodium carbonate in the arginine processing waste liquid is 25 g / L). After continuously stirring for 35 min, the solid is precipitated by standing, the solid is removed by centrifugation, and the compound amino acid content of 60 g / L is obtained by concentration; The compound amino acid content is 75.6%, and the total metal content is 16.7%; the rest are the same as in Example 1.

[0023] Control Group 1 Preparation of a mixture of the concentrated solution of the arginine processing waste liquid and the compound metal salt, including the following steps: 1) The arginine processing waste liquid is filtered through a filter membrane with a pore size of 0.45 µm to remove suspended matter, and sodium carbonate is used for the precipitation reaction. Specifically, the pH of the arginine processing waste liquid is adjusted to 8.5, and a sodium carbonate solution is added to the arginine processing waste liquid (the addition amount of sodium carbonate in the arginine processing waste liquid is 25 g / L). After continuously stirring for 35 min, the solid is precipitated by standing, the solid is removed by centrifugation, and the compound amino acid content of 60 g / L is obtained by concentration; 2) Ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate are mixed in proportion to obtain a compound metal salt, in which the molar ratio of Fe 2+ , Cu 2 + , Zn 2+ , Mn 2+ is 1:0.5:2:0.5; 3) The concentrated solution is mixed with the compound metal salt (the molar ratio of the total metal ions in the compound metal salt to the compound amino acids in the concentrated solution is 1:2.5) to obtain a mixture of the concentrated solution of the arginine processing waste liquid and the compound metal salt, and it is reserved for use.

[0024] III. Breeding experiment 3.1. Crab growth performance test Seed selection and grouping: Select the same batch of artificially hatched crab seedlings, Eriocheir sinensis ( Chinese mitten crab ), Eriocheir hepuensis ( Eriocheir hepuensis ), Eriocheir japonica ( Eriocheir japonica ), Portunus trituberculatus ( Swimming crab ), and Portunus sanguinolentus ( Portunus sanguinolentus ) with a body weight of about 50.35 g ± 2.15 g, and randomly divide them into 6 groups (Example 1 - 4 groups, Control Group 1, and Blank Group), with 4 replicates in each group and 100 in each replicate.

[0025] Feeding management: The breeding period lasts for 12 weeks. At 16:00 every day, the basic feed is fed at 4%-8% of the crab weight (6% of the crab weight is fed every day in the first 2 weeks of breeding, 8% of the crab weight is fed every day from the 3rd week to the 5th week of breeding, 6% of the crab weight is fed every day from the 6th week to the 8th week of breeding, and 4% of the crab weight is fed every day from the 9th week to the 12th week of breeding) at a fixed point. (Among them, in the first to fourth groups of Examples, the compound amino acid chelates of Examples 1-4 are incorporated into the basic feed at an incorporation amount of 2 wt%, and in Control Group 1, the concentrated solution of the arginine processing waste liquid of Control Group 1 and the mixture of complex metal salts are incorporated into the basic feed at an incorporation amount of 2 wt%; the basic feed for feeding is used as the blank group).

[0026] Test conditions: Water is changed every day during the breeding process. For Eriocheir sinensis, Eriocheir hepuensis, and Eriocheir japonica, the test water uses sand-filtered fresh lake water, and aeration is continuous during the test period. The test water temperature is 18-24 °C, and the pH is 7.2-7.5; for Portunus trituberculatus and Portunus sanguinolentus, the test water uses sand-filtered natural seawater, and aeration is continuous during the test period. The test water temperature is 22-26 °C, the pH is 7.5-7.8, and the salinity is 20-25‰.

[0027] It should be noted that on the above basis, during the test process, the feeding management methods of each group are the same. Immunization, deworming, and daily disinfection are carried out according to the conventional management measures of the farm. The composition and nutritional level of the basic feed are shown in Table 1: Table 1 Composition and nutritional level of the basic feed ; Note: The premix provides 25 mg of Fe, 20 mg of Cu, 80 mg of Zn, 26 mg of Mn, 12 mg of Mg, 0.16 mg of Se, 0.45 mg of I, and 0.6 mg of Co per kilogram of diet; 10,000 IU of vitamin A, 100 mg of vitamin E, 100 mg of vitamin C, 2,500 IU of vitamin D3, 2.2 mg of vitamin K3, 3.2 mg of vitamin B1, 15 mg of vitamin B2, 20 mg of vitamin B3, 10 mg of vitamin B4, 25 mg of vitamin B5, 0.016 mg of vitamin B12; 5 mg of folic acid, 600 mg of choline, 0.15 mg of biotin, and 200 mg of inositol.

[0028] At the end of the feeding test, the crabs are starved for 24 h, weighed, and the weight gain rate of the crabs is calculated. The weight gain rate of the crabs (WG, %) = 100×(final average weight - initial average weight) / initial average weight; the test data are recorded in Tables 2-3: Table 2 Statistical data of the growth performance - weight gain rate (WG) of Eriocheir ; Table 3 Statistical Data of Growth Performance of Swimming Crab - Weight Gain Rate (WG) Test ; Protein Efficiency Ratio (PER, %) = 100 × [Net Weight Gain of Crab / (Feed Intake × Feed Protein Content)], and the test data are recorded in Table 4 - 5 as follows: Table 4 Statistical Data of Growth Performance of Hairy Crab - Protein Efficiency Ratio (PER) Test ; Table 5 Statistical Data of Growth Performance of Swimming Crab - Protein Efficiency Ratio (PER) Test ; Experimental Conclusion: According to the data in Tables 2 - 5, it can be known that adding the compound amino acid chelate prepared by Examples 1 - 4 of the present invention to the basal diet of crabs has a certain function of promoting the growth and weight gain of crabs, and also has a certain function of improving the protein utilization efficiency of hairy crabs. Further analysis shows that the compound amino acid chelate of the present invention has a particularly significant effect on promoting the growth and weight gain of Eriocheir hepuensis and improving the protein utilization efficiency of Eriocheir hepuensis. It has a specific effect, and the precipitation reaction is increased during the pretreatment process of arginine processing waste liquid, so that the finally prepared compound amino acid chelate can further promote the growth and weight gain of Eriocheir hepuensis and significantly improve the protein utilization efficiency of Eriocheir hepuensis.

[0029] 3.2. Test on Survival Rate and Stress Response Index of Crabs under Dry Exposure Stress After the above feeding test, the crabs in Groups 1 - 4 of the examples, Control Group 1 and the blank group were subjected to dry exposure stress treatment. 100 crabs were taken from each of Groups 1 - 4 of the examples, Control Group 1 and the blank group. The specific dry exposure stress was as follows: Lay a layer of ice bags at the bottom of the foam box, and spread a layer of aquatic plants on the ice so that the crabs do not directly contact the ice. Then place the crabs neatly and without stacking in the box; lay a layer of aquatic plants on the surface of the crabs so that they do not directly contact the air. The temperature in the foam box was 18 ± 1 °C, and the relative humidity was 75 - 80%; transport for 6 h under dry exposure stress. After the transport ended, the survival rate of each group was counted and recorded in Table 6. The survival rate = (Number of Surviving Crabs / Initial Number of Crabs) × 100%; among them, death was marked by the opening of the mouth and no response when touching the four limbs.

[0030] Cortisol is one of the stress response indicator indexes. Serum cortisol was measured by radioimmunoassay (the kit was purchased from Tianjin Jiuding Medical Biotechnology Co., Ltd.). The specific measurement method was carried out according to the instructions of the kit. The test data are shown in Table 6 as follows: Table 6 Statistical Data of Survival Rate and Serum Cortisol Content of Crabs under Dry Exposure Stress ; Experimental conclusion: Based on the data analysis in Table 6, it can be known that adding the compound amino acid chelate prepared by the present invention to the basic feed of crabs can significantly reduce the serum cortisol content of Eriocheir hepuensis under the stress of exposure to air, indicating that it can alleviate the stress response of Eriocheir hepuensis. It can also significantly improve the survival rate of Eriocheir hepuensis under the stress of exposure to air, with practical application value, which can reduce the mortality rate of Eriocheir hepuensis during transportation and improve economic benefits.

[0031] The above-mentioned tests only represent several embodiments of the present invention, and their descriptions are relatively specific and detailed, but they should not be construed as limiting the scope of the present invention patent. It should be noted that for those of ordinary skill in the art, without departing from the concept of the present invention, several modifications and improvements can still be made, and these all belong to the protection scope of the present invention.

Claims

1. Application of arginine processing waste liquid in aquatic feed additive, characterized in that: The powdery compound amino acid chelate prepared by the chelation reaction of the concentrated solution after pretreatment of arginine processing waste liquid and metal salt is used as an aquatic feed additive; The pretreatment method of the arginine processing waste liquid is specifically as follows: the arginine processing waste liquid is filtered through a filter membrane to remove suspended solids, and precipitation reaction is carried out with sodium carbonate or sodium bicarbonate, and the solid is removed by centrifugation, and the concentrated solution is obtained by concentration.

2. The application of arginine processing waste liquid in aquaculture feed additives according to claim 1, characterized in that: The pore size of the filter membrane ≤ 0.45 µm.

3. The application of an arginine processing waste liquid in an aquatic feed additive according to claim 1, characterized in that: The precipitation reaction is specifically as follows: sodium carbonate or sodium bicarbonate is dissolved in deionized water to obtain a sodium carbonate solution or a sodium bicarbonate solution respectively, the pH of the arginine processing waste liquid is adjusted to 8 - 11, and the sodium carbonate solution or the sodium bicarbonate solution is added to the arginine processing waste liquid, and after continuous stirring for 30 - 40 min, the solid is precipitated by standing. Among them, the addition amount of the sodium carbonate or sodium bicarbonate in the arginine processing waste liquid is 20 - 35 g / L.

4. The application of an arginine processing waste liquid in an aquatic feed additive according to claim 1, characterized in that: The content of compound amino acids in the concentrated solution ≥ 50 g / L.

5. The application of arginine processing waste liquid in aquaculture feed additives according to claim 1, characterized in that: The chelation reaction is specifically as follows: the initial pH of the reaction solution of the chelation reaction is adjusted to 5.5 - 6.5, and under the temperature condition of 70 - 85 °C, stirring is carried out at a speed of 800 - 1500 r / min, and the chelation reaction time is 2 - 3 h.

6. The application of arginine processing waste liquid in aquatic feed additives according to claim 1, characterized in that: The metal salt is at least one of ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate.

7. Use of the arginine processing waste liquid in an aquatic feed additive according to claim 6, characterized in that: When the metal salts are a combination of ferrous sulfate, copper sulfate, zinc sulfate, and manganese sulfate, the molar ratio of Fe 2+ , Cu 2+ , Zn 2+ , Mn 2+ is 1:0.5:2:0.

5.

8. The application of an arginine processing waste liquid in an aquatic feed additive according to claim 1, characterized in that: In the preparation raw materials of the compound amino acid chelate, the molar ratio of the total amount of metal ions in the metal salt to the compound amino acids in the concentrated solution is 1:2 - 3.

9. The application of arginine processing waste liquid in aquatic feed additive according to claim 1, characterized in that: The dosage of the compound amino acid chelate as an aquatic feed additive is 0.1 wt% - 5 wt%.

10. The application of an arginine processing waste liquid in an aquatic feed additive according to claim 1, characterized in that: The compound amino acid chelate as an aquatic feed additive can promote the growth of crabs, improve the protein utilization rate of crabs, reduce the stress response of crabs under dry exposure stress, and improve the survival rate of crabs under dry exposure stress.