Functional compound feed capable of resisting ammonia nitrogen stress of micropterus salmoides and preparation method of functional compound feed

Abstract

The invention belongs to the technical field of feeds, and particularly relates to a functional compound feed capable of resisting ammonia nitrogen stress of micropterus salmoides and a preparation method of the functional compound feed. The functional compound feed comprises the following components: fish meal, cottonseed meal, chicken meal, squid paste, high gluten flour, alpha-starch, fish oil, soybean oil, soybean lecithin, monocalcium phosphate, zeolite, VC ester, a vitamin premix, a mineral premix and an amino acid mixture. The growth performance and the oxidation resistance of the largemouth bass under the ammonia nitrogen stress are improved by improving the body function of the largemouth bass; the preparation method of the functional compound feed can effectively reduce the loss of functional raw materials and improve the absorption performance of the feed.

Description

Technical Field

[0001] The invention belongs to the technical field of feed, and particularly relates to a functional compound feed resistant to ammonia nitrogen stress of largemouth bass and a preparation method thereof. Background Art

[0002] Largemouth bass (Micropterus salmoides), commonly known as California bass, belongs to the order Perciformes, family Centrarchidae, genus Micropterus. It is a warm-water carnivorous freshwater fish introduced to China from North America. California bass grows fast, has delicious meat, and few intramuscular bones, and is very popular among consumers. California bass has become an important freshwater farmed fish in my country. In 2021, the aquaculture output of California bass in my country reached 702,100 tons, second only to the output of the four major carps, crucian carp and tilapia.

[0003] In intensive aquaculture systems, the concentration of ammonia nitrogen in the water is very easy to increase due to the high density, the use of compound feeds and other factors. Therefore, ammonia nitrogen has become one of the important water quality indicators that affect the health and production of aquatic animals in addition to dissolved oxygen. With the increase in the number of largemouth bass farming, the increase in ammonia nitrogen concentration in the farming water will also cause largemouth bass health problems, resulting in economic losses. Therefore, it is urgent and important to explore functional compound feeds that can enhance the ability of largemouth bass to resist ammonia nitrogen. Summary of the invention

[0004] In view of the above problems, the object of the present invention is to provide a functional compound feed for resisting ammonia nitrogen stress of largemouth bass, and a preparation method and application thereof.

[0005] The technical content of the present invention is as follows: The present invention provides a functional compound feed for resisting ammonia nitrogen stress of largemouth bass, comprising the following components: fish meal, modified nutritional raw materials, chicken meal, squid paste, α-starch, fish oil, soybean oil, soybean lecithin, monocalcium phosphate, zeolite, VC ester, vitamin premix, mineral premix, and amino acid mixture;

[0006] The feed comprises the following components in parts by weight: 18-20 parts of fish meal, 26-32 parts of modified nutritional raw materials, 4-6 parts of chicken meal, 4-6 parts of squid paste, 4-6 parts of α-starch, 2-4 parts of fish oil, 4-6 parts of soybean oil, 0.5-1.5 parts of soybean lecithin, 0.8-3 parts of monocalcium phosphate, 3-5 parts of zeolite, 0.4-0.6 parts of VC ester, 0.4-0.6 parts of vitamin premix, 0.4-0.6 parts of mineral premix, and 16-20 parts of amino acid mixture;

[0007] The modified nutrient raw material is obtained by mixing cottonseed meal and high-gluten flour in a mass ratio of 2:1-1.4 at a solid-liquid ratio of 1:10-20 g / mL, adding 5-10 wt% urea aqueous solution, pressurizing and heating, and then centrifuging;

[0008] The pressurized heating is carried out at a pressure of 30-40 MPa and a temperature of 80-90° C. for 15-25 min;

[0009] By removing toxic substances from cottonseed meal and increasing the high-amylose content of high-gluten flour, the modified nutritional raw material is easy to digest and absorb, does not produce toxin residues after consumption, can effectively supplement protein, and increase the amount of nitrogen deposited in fish, thereby improving the disease resistance of fish;

[0010] The amino acid mixture is composed of amino acid salts, amino acid complexes and plant protein hydrolysates in a mass ratio of 1-3:10-14:11-14;

[0011] The amino acid complex includes at least three of lysine, methionine, leucine, aminobutyric acid, threonine, valine, phenylalanine, tryptophan and histidine;

[0012] The plant protein hydrolysate is obtained by homogenizing soybean meal, cooling it to 30-40° C., adding 1-6 wt % of a combined enzyme, and performing enzymolysis for 90-100 min.

[0013] The homogenization is carried out at a temperature of 80-90° C. for 20-30 minutes;

[0014] The combined enzyme consists of neutral protease, subtilisin, laccase and aspartic protease in a mass ratio of 1:0.6-0.8:1:1.

[0015] The amino acid salt is one or more of methionine salt, lysine salt and glutamate salt.

[0016] Amino acid complexes can improve the disease resistance of largemouth bass, amino acid salts can improve the dispersion of amino acids, protein hydrolysates can directly supplement a variety of amino acids, and the enzymes they contain can promote the absorption capacity of fish intestines.

[0017] The present invention also provides a method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass, comprising the following steps:

[0018] Step 1: Add soybean oil, fish oil, squid paste and soybean lecithin and heat at 60-70°C for 20-30 minutes to obtain material A;

[0019] Step 2: Mix fish meal, α-starch, chicken meal, calcium dihydrogen phosphate, zeolite and VC ester and heat to 80-90° C. for 20-30 min to obtain material B;

[0020] Step 3: Add the mineral premix and the amino acid mixture to material B and stir and keep warm for 10-20 minutes to obtain material C;

[0021] Step 4: Disperse the vitamin premix in material A, mix it evenly with the modified nutrient raw material, add it into material C, stir and keep warm for 10-20 minutes, cool and sterilize, and then dry and granulate to obtain;

[0022] The dispersion conditions are as follows: stirring in a water bath at 50-70°C in the dark for 40-60 minutes;

[0023] The process of step 4 can promote the preservation and absorption and utilization of vitamins, material A can improve the stability and dispersibility of vitamins, and the modified nutritional raw materials have the effects of encapsulation and promoting absorption.

[0024] The beneficial effects of the present invention are:

[0025] 1. The present invention provides a functional feed for largemouth bass to resist ammonia nitrogen stress. It can supplement the nutrients required by largemouth bass by compounding raw materials such as modified nutrient raw materials and amino acid mixtures, and can regulate sugar metabolism, improve body fat deposition, and maintain energy balance through signal molecules, thereby improving the growth performance and antioxidant capacity of largemouth bass under 40-day chronic ammonia nitrogen stress.

[0026] 2. The present invention provides a method for preparing a functional feed that is resistant to ammonia nitrogen stress of largemouth bass. By multiple material divisions and mixing, and temperature control, the loss of effective raw materials can be effectively reduced and the absorption performance of the feed can be improved. DETAILED DESCRIPTION

[0027] The present invention is further described in detail below through specific implementation cases. It should be understood that these embodiments are only used to illustrate the present invention and are not used to limit the scope of protection of the present invention. After reading the present invention, various equivalent forms of modifications to the present invention by those skilled in the art are all within the scope of the claims attached to this application.

[0028] Unless otherwise specified, all raw materials and reagents of the present invention are raw materials and reagents from the conventional market.

[0029] Example 1

[0030] A method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass comprises the following steps:

[0031] Step 1: Add 3 parts of fish oil, 5 parts of soybean oil, 1 part of soybean lecithin and 5 parts of squid paste, mix and heat at 65°C for 25 minutes to obtain material A;

[0032] Step 2: Mix 19 parts of fish meal, 5 parts of α-starch, 5 parts of chicken meal, 1.5 parts of monocalcium phosphate, 4 parts of zeolite and 0.5 parts of VC ester, heat to 85° C. and heat for 25 minutes to obtain material B;

[0033] Step 3: Add 0.5 parts of mineral premix and 18 parts of amino acid mixture to material B, stir and keep warm for 15 minutes to obtain material C;

[0034] Step 4: Disperse 0.5 parts of the vitamin premix in material A by stirring in a water bath at 60°C in the dark for 50 minutes, mix evenly with 29 parts of the modified nutrient raw material, add to material C, stir and keep warm for 15 minutes, cool and sterilize, and then dry and granulate to obtain;

[0035] The modified nutrient raw material is obtained by mixing cottonseed meal and high-gluten flour in a mass ratio of 2:1.2 at a solid-liquid ratio of 1:15 g / mL, adding 8wt% urea aqueous solution at 35 MPa and 85°C for 20 minutes, pressurizing and heating, and then centrifuging;

[0036] The amino acid mixture consists of methionine iron, an amino acid complex consisting of lysine, methionine, and leucine, and a plant protein hydrolysate in a mass ratio of 2:12:12;

[0037] The plant protein hydrolysate is obtained by homogenizing soybean meal at 85° C. for 25 minutes, cooling it to 35° C., adding 4 wt % of a combination enzyme consisting of neutral protease, subtilisin, laccase and aspartic protease in a mass ratio of 1:0.7:1:1, and performing enzymolysis for 95 minutes.

[0038] Example 2

[0039] A method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass comprises the following steps:

[0040] Step 1: Add 2 parts of fish oil, 4 parts of soybean oil, 0.5 parts of soybean lecithin and 4 parts of squid paste, mix and heat at 60°C for 20 minutes to obtain material A;

[0041] Step 2: Mix 18 parts of fish meal, 4 parts of α-starch, 0.8 parts of calcium dihydrogen phosphate, 4 parts of chicken meal, 0.8 parts of calcium dihydrogen phosphate, 3 parts of zeolite and 0.4 parts of VC ester, heat to 80-90° C. and heat for 20 minutes to obtain material B;

[0042] Step 3: Add 0.4 parts of mineral premix and 16 parts of amino acid mixture to material B, stir and keep warm for 10 minutes to obtain material C;

[0043] Step 4: Disperse 0.4 parts of the vitamin premix in material A by stirring in a water bath at 50°C in the dark for 40 minutes, mix evenly with 26 parts of the modified nutrient raw material, add to material C, stir and keep warm for 10 minutes, cool and sterilize, and then dry and granulate to obtain;

[0044] The modified nutrient raw material is obtained by mixing cottonseed meal and high-gluten flour in a mass ratio of 2:1 at a solid-liquid ratio of 1:10 g / mL, adding 5wt% urea aqueous solution at 30 MPa and 80° C. for 15 minutes, pressurizing and heating, and then centrifuging.

[0045] The amino acid mixture is composed of an amino acid complex consisting of lysine iron, methionine, leucine, aminobutyric acid, and threonine in a mass ratio of 1:10:11 and a plant protein hydrolysate;

[0046] The plant protein hydrolysate is obtained by homogenizing soybean meal at a temperature of 80° C. for 20 minutes, cooling it to 30° C., adding 1 wt % of a combination enzyme consisting of neutral protease, subtilisin, laccase, and aspartic protease in a mass ratio of 1:0.6:1:1, and performing enzymolysis for 90 minutes.

[0047] Example 3

[0048] A method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass comprises the following steps:

[0049] Step 1: add 4 parts of fish oil, 6 parts of soybean oil, 1.5 parts of soybean lecithin and 6 parts of squid paste, mix and heat at 70°C for 30 minutes to obtain material A;

[0050] Step 2: Mix 20 parts of fish meal, 6 parts of α-starch, 6 parts of chicken meal, 3 parts of monocalcium phosphate, 5 parts of zeolite and 0.6 parts of VC ester, heat to 90° C. and heat for 30 minutes to obtain material B;

[0051] Step 3: Add 0.6 parts of mineral premix and 20 parts of amino acid mixture to material B, stir and keep warm for 20 minutes to obtain material C;

[0052] Step 4: Disperse 0.6 parts of the vitamin premix in material A by stirring in a water bath at 70°C in the dark for 60 minutes, mix evenly with 32 parts of the modified nutrient raw material, add to material C, stir and keep warm for 20 minutes, cool and sterilize, and then dry and granulate to obtain;

[0053] The modified nutrient raw material is obtained by mixing cottonseed meal and high-gluten flour in a mass ratio of 2:1.4 at a solid-liquid ratio of 1:20 g / mL, adding 10 wt% urea aqueous solution at 40 MPa and 90° C. for 25 minutes, pressurizing and heating, and then centrifuging.

[0054] The amino acid mixture is composed of an amino acid complex consisting of zinc glutamate, aminobutyric acid, threonine, valine, phenylalanine, tryptophan and histidine in a mass ratio of 3:14:14 and a plant protein hydrolysate;

[0055] The plant protein hydrolysate is obtained by homogenizing soybean meal at 90° C. for 30 minutes, cooling it to 40° C., adding 6 wt % of a combination enzyme consisting of neutral protease, subtilisin, laccase and aspartic protease in a mass ratio of 1:0.8:1:1, and performing enzymolysis for 100 minutes.

[0056] Comparative Example 1

[0057] The difference between Comparative Example 1 and Example 1 is that no modified nutritional raw material is added in Comparative Example 1, and the other contents remain unchanged.

[0058] Comparative Example 2

[0059] The difference between Comparative Example 2 and Example 1 is that an equal amount of an amino acid complex is used in place of the amino acid mixture in Comparative Example 2, and the other contents remain unchanged.

[0060] Comparative Example 3

[0061] The difference between Comparative Example 3 and Example 1 is that the feed of Comparative Example 3 is prepared by mixing all raw materials and heating and stirring at a temperature of 70° C. for 2 hours.

[0062] First, the test of young fish largemouth bass purchased from the Jinji fish fry farm in Xiqiao Town, Nanhai District, Foshan City, Guangdong Province, after domestication for 1 week in Shunye Aquaculture Base, Shunde, Guangdong Province, 720 young fish with uniform specifications and average weight (11.18 ± 0.04g) and healthy growth were selected and randomly placed in 24 breeding barrels. The feeds prepared in the embodiments and comparative examples and the fish feeds purchased on the market were fed separately, and an ammonia nitrogen stress group was set, with 3 replicates in each group and 30 fish in each replicate. The total ammonia nitrogen concentration was set to 10% 96h-LC50, i.e. 3mg / L in ammonium chloride (NH4Cl). The breeding period was 40d. The ammonia nitrogen concentration in the water was determined by Nessler's reagent method every day in the experiment, and 10g / L NH4Cl mother solution was used to adjust the pH of the water body to be relatively stable (7.2-7.5). Feed twice a day (08:00, 17:00), feed 3-4% of fish body weight, and record sick and dead fish every day. The main water quality parameters during the breeding period are: dissolved oxygen in the water is higher than 7.0 mg / L, the average water temperature is (29±2.0)℃, and nitrite is less than 0.1 mg / L.

[0063] 2. Sample Collection and Growth Data Calculation

[0064] After the breeding experiment, the fish were fasted for 24 hours, the total weight of each bucket of fish was weighed and the number of fish was recorded, and the growth indicators such as weight gain rate and specific growth rate of tilapia were calculated. In addition, blood samples of tilapia were collected, and the physiological and biochemical enzymes and antioxidant enzyme indicators were analyzed and measured. The analysis and testing of each indicator adopted the mature methods used in domestic and foreign research. All values ​​are expressed as mean ± standard error (Mean ± SE), and each example adopts one-way analysis of variance (ANOVA) and Duncan comparison. The test results are shown in Table 1.

[0065] Table 1. Growth performance results of largemouth bass in various examples

[0066]

[0067] Note: Different letters in the same row in the table indicate significant differences (P<0.05), the same below.

[0068] It can be seen from Table 1 that the weight gain rate of the largemouth bass fed with the feed of the embodiment is increased, the feed coefficient is reduced, and the survival rate is improved; while the weight gain rate of the largemouth bass under ammonia nitrogen stress is reduced, the feed coefficient is increased, and the survival rate is reduced; the feed of the present invention can improve the ammonia nitrogen resistance of the largemouth bass.

[0069] 2. Detection of serum physiological and biochemical parameters and antioxidant enzymes of largemouth bass

[0070] The blood was collected from the tail vein of the largemouth bass and placed in a 1.5 mL centrifuge tube. After standing in a 4°C refrigerator for 2 hours, the blood was centrifuged (4°C, 4 000r / min for 10 minutes) to extract the serum, which was then stored in a -80°C refrigerator for later use. The test kits were purchased from Nanjing Jiancheng Bioengineering Institute; the test results are shown in Table 2.

[0071] Table 2 Results of serum physiological and biochemical and antioxidant enzyme index tests on largemouth bass in each case

[0072]

[0073] Note: Different letters in the same column in the table indicate significant differences (P<0.05).

[0074] As shown in Table 2, the albumin, acid phosphatase, and SOD of the largemouth bass fed with the feed in the example were significantly increased, and the MDA was significantly decreased. However, the albumin, acid phosphatase, and SOD in the serum of the largemouth bass under ammonia nitrogen stress were significantly decreased, and the MDA was significantly increased.

[0075] In summary, from Comparative Example 1, it can be seen that the present invention improves the immune performance and growth performance of largemouth seabass by modifying the nutritional raw materials to remove toxic side effects and small molecular proteins; from Comparative Example 2, it can be seen that the amino acid mixture of the present invention can improve the ammonia nitrogen resistance of largemouth seabass by effectively supplementing amino acids; from Comparative Example 3, it can be seen that the feed obtained by the preparation process of the present invention has excellent functionality.

Claims

1. A functional compound feed for resisting ammonia nitrogen stress of largemouth bass, characterized in that: Contains the following ingredients: fish meal, cottonseed meal, chicken meal, squid paste, high-gluten flour, α-starch, fish oil, soybean oil, soybean lecithin, monocalcium phosphate, zeolite, VC ester, vitamin premix, mineral premix, amino acid mixture; The amino acid mixture consists of amino acid salts, amino acid complexes and plant protein hydrolysates in a mass ratio of 1-3:10-14:11-14.

2. A functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 1, characterized in that: The invention comprises the following components in parts by weight: 18-20 parts of fish meal, 26-32 parts of modified nutritional raw materials, 4-6 parts of chicken meal, 4-6 parts of squid paste, 4-6 parts of alpha-starch, 2-4 parts of fish oil, 4-6 parts of soybean oil, 0.5-1.5 parts of soybean lecithin, 0.8-3 parts of monocalcium phosphate, 3-5 parts of zeolite, 0.4-0.6 parts of VC ester, 0.4-0.6 parts of vitamin premix, 0.4-0.6 parts of mineral premix and 16-20 parts of amino acid mixture.

3. A functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 1, characterized in that: The modified nutrient raw material is obtained by mixing cottonseed meal and high-gluten flour in a mass ratio of 2:1-1.4 at a solid-liquid ratio of 1:10-20 g / mL, adding 5-10 wt % urea aqueous solution, pressurizing and heating, and then centrifuging.

4. A functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 3, characterized in that: The pressurized heating is carried out at a pressure of 30-40 MPa and a temperature of 80-90° C. for 15-25 min.

5. The functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 1, characterized in that: The amino acid complex comprises at least three of lysine, methionine, leucine, aminobutyric acid, threonine, valine, phenylalanine, tryptophan, histidine and glycine.

6. The functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 1, characterized in that: The plant protein hydrolysate is obtained by homogenizing soybean meal, cooling it to 30-40°C, adding 1-6wt% of combined enzymes and performing enzymolysis for 90-100 minutes; the homogenization is performed at a temperature of 80-90°C for 20-30 minutes.

7. A functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 6, characterized in that: The combined enzyme consists of neutral protease, subtilisin, laccase and aspartic protease in a mass ratio of 1:0.6-0.8:1:

1.

8. The functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 1, characterized in that: The amino acid salt is one or more of methionine salt, lysine salt and glutamate salt.

9. The method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to any one of claims 1 to 8, characterized in that: The following steps are involved: Step 1: Add soybean oil, fish oil, squid paste and soybean lecithin and heat at 60-70°C for 20-30 minutes to obtain material A; Step 2: Mix fish meal, α-starch, chicken meal, calcium dihydrogen phosphate, zeolite and VC ester and heat to 80-90° C. for 20-30 min to obtain material B; Step 3: Add the mineral premix and the amino acid mixture to material B and stir and keep warm for 10-20 minutes to obtain material C; Step 4: Disperse the vitamin premix in material A, mix it evenly with the modified nutritional raw material, add it into material C, stir and keep warm for 10-20 minutes, cool and sterilize, and then dry and granulate to obtain.

10. The method for preparing a functional compound feed for resisting ammonia nitrogen stress of largemouth bass according to claim 9, characterized in that: The dispersion conditions in step 4 are stirring in a water bath at 50-70° C. in the dark for 40-60 min.

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