CN112515054A - Low-protein feed for improving growth performance of litopenaeus vannamei and application thereof - Google Patents
Low-protein feed for improving growth performance of litopenaeus vannamei and application thereof Download PDFInfo
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- CN112515054A CN112515054A CN202011231439.7A CN202011231439A CN112515054A CN 112515054 A CN112515054 A CN 112515054A CN 202011231439 A CN202011231439 A CN 202011231439A CN 112515054 A CN112515054 A CN 112515054A
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- improving
- low
- litopenaeus vannamei
- protein
- feed
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Classifications
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A—HUMAN NECESSITIES
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- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
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Abstract
The invention belongs to the technical field of aquaculture, and particularly relates to a low-protein feed for improving the growth performance of litopenaeus vannamei and application thereof. The low-protein feed comprises nucleotide, and is used for improving the digestion and absorption capacity of litopenaeus vannamei on feed nutrient substances, improving the growth performance of the litopenaeus vannamei, improving the shrimp body composition and serum biochemical indexes, and improving the immunity. The nucleotide comprises 5 '-cytidylic acid, 5' -adenylic acid, 5 '-inosinic acid disodium, 5' -uridylic acid disodium, 5 '-guanylic acid disodium and 5' -thymidylic acid disodium, and is mainly used for improving the crude fat content of litopenaeus vannamei, serum glutamic-oxalacetic transaminase, glutamic-pyruvic transaminase, hepatopancreatic glutamic-pyruvic transaminase, blood cell number, serum PO activity, and total protein content of stomach and intestinal tracts.
Description
Technical Field
The invention belongs to the technical field of aquaculture, and particularly relates to a low-protein feed for improving the growth performance of litopenaeus vannamei and application thereof.
Background
China is a big aquaculture country, and the aquaculture yield is the first world for years. Feed protein is the main energy source substance of aquatic animals. Protein resources (such as fish meal) used in the aquatic feed industry of China are in a shortage state for a long time, and the dependence import degree is high, so that the sustainable development of the aquatic feed industry and the aquaculture industry of China is restricted. However, with the rapid development of aquaculture in China, the industrial mode gradually changes to high-density intensive culture, and the waste water in the culture process is increased, so that water pollution is easily caused, the ecological environment is destroyed, and the ecological culture construction is not facilitated. Energy conservation and emission reduction are always the direction of concern of the aquaculture industry. The technology of low-protein feed (also called balanced amino acid feed) has the advantages of reducing the addition of protein raw materials, reducing nitrogen emission and even improving the growth performance of animals, and is an effective means for energy conservation and emission reduction.
The low protein feed technology has been deeply researched in livestock breeding, particularly pig production, and further researched in poultry. In aquaculture, low protein feed technology is developed to a certain extent, and the application of the low protein feed technology in aquaculture needs further research.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a low-protein feed for improving the growth performance of litopenaeus vannamei and application thereof, and provides a low-protein feed for improving the digestion and absorption capacity of the litopenaeus vannamei on feed nutrients, improving the shrimp body composition and serum biochemical indexes and improving the immunity.
The technical content of the invention is as follows:
the invention provides an application of a low-protein feed for improving the growth performance of Litopenaeus vannamei, wherein the low-protein feed is used for improving the digestion and absorption capacity of the Litopenaeus vannamei on feed nutrients, improving the shrimp body composition and serum biochemical indexes, and improving the immunity;
the low-protein feed is used for improving the weight gain rate of the litopenaeus vannamei, the content of body crude fat and ash, the activity of serum glutamic-oxaloacetic transaminase, the activity of glutamic-pyruvic transaminase, the content of uric acid, the activity of hepatopancreatic glutamic-pyruvic transaminase, the number of blood cells and the content of total protein in stomach and intestinal tracts;
the low-protein feed is used for improving the growth performance, body composition, serum biochemistry and immune antioxidation of the litopenaeus vannamei.
The invention also provides a low-protein feed for improving the growth performance of the litopenaeus vannamei, which comprises nucleotide, fish meal, cuttlefish paste, shrimp shell powder, soybean protein concentrate, blood meal, peanut bran, soybean meal, flour, fish oil, soybean phospholipid oil, cholesterol, 35% VC ester, monocalcium phosphate, zeolite powder, choline chloride, vitamin premix, mineral premix, sodium carboxymethylcellulose, cellulose and crystal methionine;
the nucleotide comprises 15-25% of 5 '-cytidylic acid, 15-25% of 5' -adenylic acid, 15-25% of disodium 5 '-inosinate, 15-25% of disodium 5' -uridylate, 15-25% of disodium 5 '-guanylate and 15-25% of disodium 5' -thymidylate;
the content of protein in the low-protein feed is 30-40%.
The invention has the following beneficial effects:
compared with high-protein and low-protein level feeds, the low-protein feed for improving the growth performance of the litopenaeus vannamei contains nucleotide raw materials, and the nucleotide mixture added into the low-protein feed has the advantages that the capability of the litopenaeus vannamei on digestion and absorption of substances is improved, and the growth condition of the litopenaeus vannamei is promoted;
the low-protein feed containing nucleotide has the effect of improving the growth performance, the feed utilization efficiency, the shrimp body composition, the serum biochemical index and the immunity of the litopenaeus vannamei, and particularly has obvious effects on the crude fat content of the litopenaeus vannamei, serum glutamic-oxaloacetic transaminase, serum glutamic-pyruvic transaminase, hepatopancreatic glutamic-pyruvic transaminase, the blood cell number, the serum PO activity and the total protein content of the stomach and intestinal tracts.
Detailed Description
The present invention is described in further detail in the following detailed description with reference to specific embodiments, which are intended to be illustrative only and not to be limiting of the scope of the invention, as various equivalent modifications of the invention will become apparent to those skilled in the art after reading the present invention and are intended to be included within the scope of the appended claims.
All the raw materials and reagents of the invention are conventional market raw materials and reagents unless otherwise specified.
Examples
A low-protein feed for improving the growth performance of Litopenaeus vannamei comprises:
1) the low-protein feed comprises the following components: the feed additive is prepared by mixing nucleotide (5 '-cytidylic acid, 5' -adenylic acid, 5 '-inosinic acid disodium, 5' -uridylic acid disodium, 5 '-guanylic acid disodium and 5' -thymidylic acid disodium in a mass ratio of 1:1:1:1:1:1, wherein the proportion can be adjusted according to actual conditions, the usage amount of each nucleotide is controlled to be 15-25%), fish meal, cuttlefish cream, shrimp shell powder, soybean protein concentrate, blood meal, peanut bran, soybean meal, flour, fish oil, soybean phospholipid oil, cholesterol, 35% VC ester, calcium dihydrogen phosphate, zeolite powder, 50% choline chloride, vitamin premix, mineral premix, sodium carboxymethylcellulose, cellulose and crystal methionine;
in the embodiment, the low-protein group feed and the high-protein group feed are respectively provided with an experimental group and a control group (4 groups in total), the experimental group is prepared by adding nucleotide mixture on the basis of the feed of the control group, and the feed of the 4 groups is prepared in an equal-fat mode;
pulverizing all feed materials, sieving with 80 mesh sieve, mixing trace components by stepwise expansion method, adding oil and water, making into granule with diameter of 2.0mm with double screw press (SLX-80 type), oven drying at 55 deg.C for 7-8 hr, naturally cooling, sieving, bagging, and storing at room temperature;
the feed formulation for each test group is shown in table 1 and embodies nutritional levels, amino acid and fatty acid composition.
TABLE 1 prawn feed base formula composition and nutrient level (air-dry base)
Wherein, the vitamin premix provides VA 8000IU, VD 34000 IU, VE 60mg, VK 320 mg, VB 110 mg, VB 230 mg, VB 616 mg, 50mg of calcium pantothenate, 5mg of folic acid, 0.16mg of biotin, 60.3mg of nicotinic acid, VB120.03mg and 300mg of inositol for each kilogram of feed;
the mineral premix provides MgSO for each kilogram of feed4·H2O 60mg、KCl 450mg、Met-Cu 15mg、FeSO4·H2O 5mg、ZnSO4·H2O 50mg、Ca(IO3)2 0.3mg、Met-Co 0.8mg、NaSeO3 0.018mg。
2) Litopenaeus vannamei breeding
The Litopenaeus vannamei purchased from the unified shrimp farm in Zhongshan City of Guangdong province is temporarily raised for 2 weeks in an indoor circulating water culture system by using commercial feed, and fed for 3 times every day at 8:00, 14:00 and 20: 00;
the indoor circulating water culture system is composed of a glass fiber barrel;
480 litopenaeus vannamei fries with healthy physique and 1.0g of individual weight are selected and randomly divided into 4 groups, and each group is fed with 4 feeds in the table 1.
Feeding for three times at 8:30, 14:30 and 20:30 every day by adopting a satiation feeding method, wherein the feeding amount is 4-6% of the body weight, sucking out residual bait 30min after feeding, and meanwhile, timely adjusting the feeding amount according to the ingestion condition.
The method comprises the following steps of (1) feeding litopenaeus vannamei in a saline-fresh water environment formed by mixing seawater and tap water, sand filtering, disinfecting and adding, wherein the salinity is 4.0-5.0 per mill, natural illumination is performed, the water temperature is 20-26 ℃, the pH value is 7.0-8.0, the dissolved oxygen is greater than 6.0mg/L, the ammonia nitrogen concentration is less than or equal to 0.1mg/L, and the nitrite concentration is less than or equal to 0.01 mg/L;
the water quality was measured every week during the rearing period 56 d.
After the feeding period is finished, fasting is carried out for 12 hours, the growth condition, the body type index, the feed utilization condition, the shrimp body components, the shrimp serum biochemical index, the shrimp tissue enzyme activity, the shrimp immunity index and other results of the litopenaeus vannamei are detected and result analysis is carried out, the experimental result is represented by the Mean value plus or minus standard deviation (Mean plus or minus SD), two-factor variance analysis is carried out by Spss17.0 software, and significant difference analysis is carried out among Duncan's multiple comparison groups, wherein the significant difference level is P < 0.05.
1.2 Effect of nucleotide addition to feed containing protein level on Litopenaeus vannamei growth Performance
Weighing terminal weight, counting survival mantissas of the prawns, and calculating Weight Gain Rate (WGR), Specific Growth Rate (SGR), Food Intake (FI), Feed Coefficient (FC) and Survival Rate (SR) of the prawns, wherein the results are shown in the following table:
TABLE 2 Litopenaeus vannamei growth Performance Condition
Wherein, the weight gain ratio (WGR,%) is (final weight-initial weight) × 100/initial weight;
specific growth rate (SGR,%/d) ([ ln (last average weight) — ln (first average weight) ] × 100/feeding day;
survival (SR,%) × 100 (shrimp tail at end of experiment/shrimp tail at start of experiment);
the feed Factor (FC) is the feed amount/(final shrimp weight + dead shrimp weight in test-initial shrimp weight).
As can be seen from Table 2, the low protein level in the feed and the addition of the mixed nucleotide based on the low protein level can improve the final average weight, the survival rate, the weight gain rate and the specific growth rate of the litopenaeus vannamei, reduce the feed coefficient, and obviously influence the final average weight, the weight gain rate and the specific growth rate of the litopenaeus vannamei by the protein level and the nucleotide level in the feed.
2.2 Effect of nucleotide addition to feed at protein level on Litopenaeus vannamei form index and feed utilization
TABLE 3 Litopenaeus vannamei body index and feed utilization
Wherein, hepato-pancreatic index (HSI,%) (weight of hepato-pancreas/body weight) × 100;
fullness (CF, g/cm)3) 100 x weight of each shrimp/(length of each shrimp)3；
Feed intake (FI, g/shrimp) ═ total bait feed/[ (shrimp tail at the beginning of the test + shrimp tail at the end of the test)/2 ];
protein retention (PDR,%) 100 × (final shrimp body protein assay-initial shrimp body protein assay)/protein intake.
As shown in Table 3, the low protein level can significantly improve the food intake and protein retention of the prawns, and the nucleotide group can improve the food intake and protein retention of the prawns.
3.2 Effect of nucleotide addition to feed at protein level on the constituents of Litopenaeus vannamei
Tail shrimp were selected for each group for determination of whole shrimp dry matter, crude protein, crude fat and ash, with the results shown in the following table:
TABLE 4 Litopenaeus vannamei body composition (dry weight,%)
As can be seen from table 4, the protein level in the feed significantly affected the shrimp body crude fat content, which was significantly lower in the low protein group than in the high protein group. The addition of nucleotide in the feed can improve the content of crude protein of the shrimp bodies, and obviously improve the content of dry substances, crude fat and ash in the shrimp bodies. Protein levels and nucleotide levels have an interaction on shrimp body crude fat content.
4.2 Effect of nucleotide added into feed with protein level on biochemical indexes of blood serum of Litopenaeus vannamei
Shrimp were taken from each group and serum was prepared: taking blood from a pericardial cavity by using a 1mL sterile syringe, combining the blood and placing the blood in a sterile Eppendorf tube, standing the blood in a refrigerator at 4 ℃ overnight, centrifuging the blood at 10000r/min for 10min, removing serum for subpackaging, storing the blood in the refrigerator at-80 ℃ for later use, and detecting biochemical indexes in the serum;
the measurement of Total Protein (TP), Lysozyme (LZM), alkaline phosphatase (AKP), glutamic-oxalacetic transaminase (AST) and glutamic-pyruvic transaminase (ALT) of the homogenate supernatant is carried out by adopting a kit instruction corresponding to the Nanjing construction kit;
the results are shown in the following table:
TABLE 5 Biochemical index of Litopenaeus vannamei serum
As can be seen from Table 5, the low protein group significantly increased the cholesterol level in the shrimp serum and significantly decreased the glutamate pyruvate transaminase activity. The protein level has no significant effect on glutamic-oxaloacetic transaminase, uric acid, blood sugar and total protein content in the blood serum of the shrimps. The addition of nucleotide in the feed can obviously reduce the activity of glutamic-oxalacetic transaminase and glutamic-pyruvic transaminase in the blood serum of prawns, reduce the total protein content in the blood serum, and has no obvious influence on the contents of cholesterol, uric acid and blood sugar. There is an interaction between protein levels and nucleotide levels in the activity of aspartate aminotransferase and glutamate pyruvate transaminase.
5.2 Effect of nucleotide addition to feed at protein level on AKP, AST and ALT vitality of Litopenaeus vannamei tissues
Taking 15 shrimps from each of the 4 groups of the litopenaeus vannamei, peeling off hepatopancreas, stomachs and intestinal tracts, and storing in a refrigerator at the temperature of-80 ℃ for later use;
preparation of homogenates of the liver, pancreas, stomach and intestinal tissues supernatants: taking a hepatopancreas, stomach or intestinal tract sample out of a refrigerator at the temperature of-80 ℃ or 4 ℃, unfreezing at the temperature of 4 ℃, then weighing about 0.3g of a sheared sample, putting the sheared sample into a glass homogenizer, adding precooled physiological saline with the weight 6 times that of the sample, homogenizing in ice bath (dY 89-II type electric glass homogenizer, Ningbo New Ganoderma biotech Ltd.), pouring the homogenate into a centrifuge tube after sufficient homogenization, adding precooled physiological saline with the weight 3 times that of the sample, washing the homogenizer, pouring into the centrifuge tube to prepare 10% tissue homogenate, centrifuging (eppendorf centrifuge 5804R) for 10min at the temperature of 4 ℃ at 3000R/min, taking supernatant, subpackaging and storing in the refrigerator at the temperature of-80 ℃ for later use.
Phenol Oxidase (PO) assay: reference is made to the method of Ashida (1971), with minor modifications. Using L-dopa as a substrate, 10. mu.L of serum (tissue homogenate) was added to a 96-well microplate, 200. mu.L of phosphate buffer solution with a concentration of 0.1mol/L, pH of 6.0 was then added to each well, and finally 10. mu.L of L-dopa solution with a concentration of 0.01mol/L (purchased from sigma) was added to each sample well, and the mixture was shaken 4 times in an microplate reader (550, Bio-Rad), and the absorbance at 490nm was read every 4 min. The enzyme activity is 1 enzyme activity unit by increasing 0.001 per minute of OD490 value under the test condition.
The measurement results of the above-described indices are shown in tables 6 and 7:
TABLE 6 Litopenaeus vannamei AKP, AST and ALT vitality
As can be seen from table 6, protein levels in the feed significantly affected the total protein content of the hepatopancreas, stomach and intestinal tract. The low protein level significantly increases the total protein content of the intestinal tract, and significantly reduces the total protein content of the liver, pancreas and stomach. The addition of nucleotide in the feed can obviously improve the quantity of prawn blood cells, the activity of serum PO and the content of intestinal TP. Protein and nucleotide levels have significant interactions in blood cell number, serum PO viability, total gastric and intestinal protein content.
TABLE 7 immunological index profiles of Litopenaeus vannamei
As can be seen from table 7, protein levels in the feed significantly affected the total protein content of the hepatopancreas, stomach and intestinal tract. The low protein level significantly increases the total protein content of the intestinal tract, and significantly reduces the total protein content of the liver, pancreas and stomach. The addition of nucleotide in the feed can obviously improve the quantity of prawn blood cells, the activity of serum PO and the content of intestinal TP. Protein and nucleotide levels have significant interactions in blood cell number, serum PO viability, total gastric and intestinal protein content.
Therefore, the addition of nucleotide in the low protein level can obviously improve the growth performance of the prawns, improve the utilization efficiency of the feed, improve the composition of the prawns and the biochemical indexes of serum, and improve the immunity. Protein levels and nucleotide addition levels have significant interactions in shrimp body crude fat content, serum glutamic-oxaloacetic transaminase and glutamic-pyruvic transaminase, hepatopancreatic glutamic-pyruvic transaminase, blood cell count, serum PO activity, and total protein content in stomach and intestinal tracts.
In conclusion, the nucleotide-containing low-protein feed has an improvement effect on the growth performance, the feed utilization efficiency, the shrimp body composition, the serum biochemical indexes and the immunity of the litopenaeus vannamei, and particularly has an obvious effect on the crude fat content of the litopenaeus vannamei, serum glutamic-oxaloacetic transaminase, glutamic-pyruvic transaminase, hepatopancreatic glutamic-pyruvic transaminase, the number of blood cells, the serum PO activity and the total protein content of the stomach and intestinal tracts.
Claims (6)
1. The application of the low-protein feed for improving the growth performance of the litopenaeus vannamei is characterized in that the components of the low-protein feed comprise nucleotides, and the low-protein feed is used for improving the digestion and absorption capacity of the litopenaeus vannamei on nutrient substances of the feed, improving the growth performance, improving the shrimp body composition and the serum biochemical index and improving the immunity.
2. The use of the low-protein feed for improving the growth performance of litopenaeus vannamei according to claim 1, wherein the low-protein feed is used for improving the weight gain rate of litopenaeus vannamei, the content of body fat and ash, the activity of serum glutamic-oxaloacetic transaminase, the activity of glutamic-pyruvic transaminase and uric acid, the activity of hepatopancreatic glutamic-pyruvic transaminase, the number of blood cells, and the content of total protein in stomach and intestinal tracts.
3. The low-protein feed for improving the growth performance of the litopenaeus vannamei is characterized by comprising nucleotide, fish meal, cuttlefish paste, shrimp shell powder, soybean protein concentrate, blood meal, peanut bran, soybean meal, flour, fish oil, soybean phospholipid oil, cholesterol, 35% VC ester, monocalcium phosphate, zeolite powder, choline chloride, vitamin premix, mineral premix, sodium carboxymethylcellulose, cellulose and crystal methionine.
4. The low-protein feed for improving the growth performance of litopenaeus vannamei as claimed in claim 3, wherein the nucleotides comprise 5 '-cytidylic acid, 5' -adenylic acid, disodium 5 '-inosinate, disodium 5' -uridylic acid, disodium 5 '-guanylic acid and disodium 5' -thymidylic acid.
5. The low-protein feed for improving the growth performance of Litopenaeus vannamei Boone as claimed in claim 4, wherein the nucleotide comprises 15-25% of 5 '-cytidylic acid, 15-25% of 5' -adenylic acid, 15-25% of disodium 5 '-inosinate, 15-25% of disodium 5' -uridylate, 15-25% of disodium 5 '-guanylate and 15-25% of disodium 5' -thymidylate.
6. The low-protein feed for improving the growth performance of litopenaeus vannamei according to claim 3, wherein the content of protein in the low-protein feed is 30-40%.
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Application publication date: 20210319 |