Shrimp larva feed as well as preparation method and application thereof

Abstract

The invention discloses shrimp seed feed as well as a preparation method and application thereof. Belongs to the technical field of aquatic product feed preparation. According to the application, the aspergillus niger fermented peanut cake (FGNC) and the food residue fermented protein (FKR) are compounded to replace the fish meal, the problem of amino acid imbalance of single plant protein is solved through a protein complementary effect, the replacement rate of the fish meal reaches 70% through compounding of the aspergillus niger fermented peanut cake and the food residue fermented protein, and growth inhibition is avoided; meanwhile, lactic acid in the FKR can activate phytase secreted by aspergillus niger in the FGNC accidentally, the degradation rate of the phytic acid is increased, the problem of vegetable protein mineral absorption obstacle is thoroughly solved, and technical support is provided for preparation of low-cost and high-efficacy shrimp seed feed.

Description

Technical Field

[0001] This invention relates to the field of aquatic product feed preparation technology, and more specifically to a shrimp larvae feed, its preparation method, and its application. Background Technology

[0002] Shrimp larvae feed is a specialized aquatic feed designed specifically for the larval stage of shrimp (the transition period from hatching to juvenile shrimp). Its core function is to provide balanced nutrition for the larvae, supporting shell formation, digestive system development, and immune system establishment, directly affecting the survival rate, growth rate, and disease resistance of the larvae. It is mainly divided into: Initial feeding: for larvae aged 0-15 days after hatching, with a particle size ≤200μm, requiring high protein, easy digestibility, and immune-enhancing components. Stage-specific feeding: the formula is dynamically adjusted according to the larval development cycle; for example, high cholesterol and minerals are needed during molting, while muscle growth is emphasized during the rearing period.

[0003] Shrimp larvae feed technology faces multiple challenges: First, fishmeal substitution technology has not yet been mastered. Traditional feeds contain 50-70% fishmeal, leading to high costs and a growing shortage of fishmeal resources. Furthermore, replacing fishmeal with plant protein exceeding a certain proportion (30%) results in decreased growth and amino acid imbalance. Second, the effects of functional additives are unstable. Active ingredients such as astaxanthin and Bacillus subtilis are easily inactivated during high-temperature pelleting, and conventional microencapsulation technology is costly. Third, environmental pollutant emissions are severe. Traditional feeds have a nitrogen utilization rate of only 25%, and uneaten feed contributes to eutrophication of water bodies. In addition, existing feeds lack precise nutritional regulation and cannot match the dynamic nutritional needs of shrimp larvae during molting and rearing stages.

[0004] Therefore, how to solve the technical problems existing in current shrimp larvae feed, such as the difficulty in replacing fishmeal, the inactivation of functional additives, and the high emissions of environmental pollutants, and how to significantly improve the growth performance and health of shrimp larvae through innovative raw material combinations and preparation processes, is a technical problem that urgently needs to be solved by those skilled in the art. Summary of the Invention

[0005] In view of this, the present invention provides a shrimp larvae feed, a preparation method thereof and its application. The present invention solves the technical bottlenecks in existing shrimp larvae feeds, such as the difficulty in replacing fishmeal, the inactivation of functional additives, and the high emissions of environmental pollutants.

[0006] To achieve the above objectives, this application adopts the following technical solution:

[0007] A type of shrimp larvae feed comprises the following components in parts by weight: 20-25 parts of Aspergillus niger fermented peanut cake, 15-20 parts of meal waste fermented protein, 10-15 parts of Antarctic krill powder, 0.5-1.2 parts of double-encapsulated astaxanthin microspheres, 0.8-1.5 parts of double-encapsulated Bacillus licheniformis, 1-2 parts of seaweed polysaccharide, 2-3 parts of phospholipid oil DHA, and 3-6 parts of dynamic nutrient components.

[0008] By using a combination of Aspergillus niger fermented peanut cake (FGNC) and food waste fermentation products (FKR) to replace fish meal, the problem of amino acid imbalance in single plant proteins can be solved through protein complementarity.

[0009] As a preferred technical solution, when shrimp larvae are in the molting stage, the dynamic nutrient component is a high-cholesterol concentrate; when shrimp larvae are in the growth stage, the dynamic nutrient component is a high-peptide protein. Based on the different needs of shrimp larvae during the molting stage (high cholesterol) and the growth stage (high protein), dynamic nutrient components are designed to achieve precise nutrition at each stage.

[0010] Another object of this application is to provide a method for preparing the shrimp larvae feed, comprising the following steps:

[0011] (1) Preparation of peanut cake fermented with Aspergillus niger: Peanut cake was crushed, the moisture content was adjusted to 48-50%, and it was steamed at 121℃ for 15-20 minutes. Then, it was inoculated with Aspergillus niger for solid-state fermentation to prepare peanut cake fermented with Aspergillus niger. Aspergillus niger (AS3.350) secretes phytase and neutral protease. Phytase can degrade phytic acid and release calcium, zinc, iron and other minerals chelated by phytic acid, thereby improving bioavailability. Neutral protease can increase the content of small molecule protein peptides in the raw materials, thereby improving the digestibility of feed for shrimp larvae. In addition, peanut cake fermented with Aspergillus niger can also eliminate anti-nutritional factors such as trypsin inhibitors and tannins, reducing their content.

[0012] (2) Preparation of fermented protein from food waste: After sorting and crushing food waste, inoculate it with compound microbial agent and ferment it in solid state at 36-39℃ for 36-48h to prepare fermented protein from food waste.

[0013] (3) Preparation of double-encapsulated astaxanthin microspheres and double-encapsulated Bacillus licheniformis: Mix astaxanthin or Bacillus licheniformis powder with sodium alginate solution, add CaCl2 solution dropwise, let stand for 10-15 min, and solidify to form gel microspheres; immerse the gel microspheres in 1.0-1.5% chitosan acetate solution, adjust the pH to 5.8, stir for 25-35 min, take out the gel microspheres, drain, pre-freeze at -40℃ and vacuum dry to obtain double-encapsulated astaxanthin microspheres or double-encapsulated Bacillus licheniformis; use sodium alginate-chitosan double encapsulation process to protect heat-sensitive components such as astaxanthin and Bacillus licheniformis to achieve targeted release in the intestine.

[0014] (4) Mix Aspergillus niger fermented peanut cake with meal waste fermented protein, add papain, and hydrolyze at 45-55℃ for 2 hours; then add phospholipid oil DHA and dynamic nutrient components, and homogenize at high speed to form an O / W emulsion; then add Antarctic krill powder, double-encapsulated astaxanthin microspheres, double-encapsulated Bacillus licheniformis and seaweed polysaccharide, mix well, quick-freeze at -80℃ and then freeze-dry under vacuum, crush and sieve to obtain 200-500μm particles, which is shrimp larvae feed. Aspergillus niger fermented peanut cake (FGNC) and meal waste fermented protein (FKR) are combined. The lactic acid in FKR can activate the Aspergillus niger phytase remaining in FGNC, further improving the phytic acid degradation rate compared with FGNC alone; and the two also have a complementary nutritional effect.

[0015] FGNC lacks methionine (a sulfur-containing amino acid), while the sulfur-containing amino acids (methionine + cysteine) in FKR directly fill the methionine gap in FGNC.

[0016] Butyric acid in FKR short-chain fatty acids (SCFAs) can activate HDAC enzymes in intestinal cells, upregulate the expression of transport protein PepT1, and thus promote the absorption of FGNC small peptides.

[0017] The three-step low-temperature preparation process replaces the traditional high-temperature granulation with enzymatic hydrolysis, emulsification, and vacuum freeze-drying, thus preserving the biological activity completely.

[0018] As a preferred technical solution, in step (1), the peanut cake is crushed to ≤2mm; the concentration of Aspergillus niger is 10. 6 The inoculum concentration was CFU / mL, and the inoculum size was 1-1.5% v / w. During the solid-state fermentation process, 0.2% KH2PO4 by mass was also added. The solid-state fermentation temperature was 30±1℃, the fermentation time was 35-40h, and the stirring frequency was once every 12-16h.

[0019] As a preferred technical solution, the compound microbial agent in step (2) is a compound microbial agent of Lactobacillus plantarum and Bacillus subtilis, with a ratio of Lactobacillus plantarum to Bacillus subtilis ≥ 3:1; the amount of the compound microbial agent added is 3-5% of the amount of food waste. When fermentation is carried out using the compound microbial agent, lactic acid and short-chain fatty acids (acetic acid, propionic acid and butyric acid) will be produced in the metabolic products. Lactic acid products can reduce the pH value of the intestine, thereby inhibiting the proliferation of Vibrio; while butyric acid in the short-chain fatty acids can promote the repair of intestinal epithelial cells. In addition, the proportion of Lactobacillus plantarum > 75% ensures that acid production is dominant and inhibits the growth of Vibrio, while Bacillus subtilis ≤ 25% avoids excessive amine production (amines > 200 mg / kg inhibit growth).

[0020] As a preferred technical solution, the amount of sodium alginate solution added in step (3) is 3-4% of the mass of astaxanthin or Bacillus licheniformis powder; the amount of CaCl2 solution added is 1.5-2% of the mass of astaxanthin or Bacillus licheniformis powder.

[0021] As a preferred technical solution, the amount of papain added in step (4) is 0.2-0.5% of the total mass of peanut cake fermented with Aspergillus niger and fermented protein from leftover food; the speed of high-speed homogenization in step (4) is 10000-15000 rpm and the time is 8-15 min.

[0022] As a preferred technical solution, when the shrimp larvae are in the molting period, the dynamic nutrient component is a high cholesterol concentrate, which is prepared by the following method: shrimp shell powder and squid liver enzymatic hydrolysate are mixed and emulsified at a weight ratio of 2:1.

[0023] As a preferred technical solution, when the shrimp larvae are in the growth period, the dynamic nutrient component is a high peptide protein, which is prepared by the following method: fish lysate is prepared by enzymatic hydrolysis with neutral protease at 42-46℃ for 3-5 hours.

[0024] Another object of this application is to provide the application of the shrimp larvae feed described herein or the shrimp larvae feed prepared by the method described herein in the rearing process of Litopenaeus vannamei.

[0025] Another object of this application is to provide the application of the combination of Aspergillus niger fermented peanut cake and meal waste fermented protein prepared in steps (1) and (2) above as a fish meal substitute in the preparation of shrimp larvae feed.

[0026] As can be seen from the above technical solution, compared with the prior art, the present invention has the following beneficial effects:

[0027] (1) This application uses Aspergillus niger fermented peanut cake (FGNC) and food waste fermentation product (FKR) to replace fish meal. The protein complementarity effect solves the problem of amino acid imbalance of single plant protein. The combination of the two makes the fish meal replacement rate reach 70%, and there is no growth inhibition, improves nitrogen utilization, and solves the problem of environmental pollution emissions.

[0028] (2) Double encapsulation active protection technology: Sodium alginate-chitosan double encapsulation process is used to protect heat-sensitive components such as astaxanthin and Bacillus subtilis, so as to achieve targeted release in the intestine;

[0029] (3) Dynamic nutrition regulation technology: Based on the molting cycle of shrimp larvae, cholesterol and protein gradients are designed to achieve precise nutrition at each stage;

[0030] (4) Three-step low-temperature preparation process: enzymatic hydrolysis → emulsification → vacuum freeze drying replaces traditional high-temperature granulation, thus fully preserving biological activity. Detailed Implementation

[0031] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0032] Example 1

[0033] A shrimp larvae feed comprises the following components in parts by weight:

[0034] 20 portions of Aspergillus niger fermented peanut cake, 15 portions of meal waste fermented protein, 10 portions of Antarctic krill powder, 0.5 portions of double-encapsulated astaxanthin microspheres, 0.8 portions of double-encapsulated Bacillus licheniformis, 1 portion of seaweed polysaccharide, 2 portions of phospholipid oil DHA, and 3 portions of dynamic nutrient components;

[0035] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0036] The specific preparation method of the above-mentioned shrimp larvae feed is as follows:

[0037] (1) Preparation of peanut cake fermented with Aspergillus niger: The peanut cake was crushed to ≤2mm, the moisture content was adjusted to 48%, and it was steamed at 121℃ for 15min. Then, it was inoculated with Aspergillus niger (Aspergillus niger AS3.350, concentration 10). 6 CFU / mL, inoculation amount of 1% v / w (commercially available), and 0.2% KH2PO4 were added. Solid fermentation was carried out at 30±1℃ for 35h, with stirring once every 12h to prepare Aspergillus niger fermented peanut cake.

[0038] (2) Preparation of fermented protein from food waste: After sorting and crushing the food waste, inoculate it with a compound bacterial agent of Lactobacillus plantarum and Bacillus subtilis (Lactobacillus plantarum: Bacillus subtilis ≥ 3:1, with an addition amount of 3%), and ferment it in solid state at 36℃ for 36h to obtain fermented protein from food waste.

[0039] (3) Preparation of double-encapsulated astaxanthin microspheres and double-encapsulated Bacillus licheniformis: Mix astaxanthin or Bacillus licheniformis powder with sodium alginate solution of 3% by weight of astaxanthin or Bacillus licheniformis powder, add 1.5% by weight of CaCl2 solution of astaxanthin or Bacillus licheniformis powder, let stand for 10 min, solidify to form gel microspheres; immerse the gel microspheres in 1.0% chitosan acetate solution, adjust the pH to 5.8, stir for 25 min, take out the gel microspheres, drain, pre-freeze at -40℃ and vacuum dry to obtain double-encapsulated astaxanthin microspheres or double-encapsulated Bacillus licheniformis;

[0040] (4) Preparation of high cholesterol concentrate: Cuttlefish liver was enzymatically hydrolyzed with neutral protease (addition amount 0.5%) at 45℃ for 4 hours to release bound cholesterol and prepare cuttlefish liver enzymatic hydrolysate; the prepared cuttlefish liver enzymatic hydrolysate was mixed with shrimp shell powder (300 mesh) at a ratio of 1:2, and 5% by volume of phospholipid emulsifier (soy lecithin) was added to the mixture. The mixture was homogenized at 15000 rpm for 10 minutes to prepare high cholesterol concentrate;

[0041] Preparation of high-peptide protein: Fish processing by-products (fish head, viscera, and minced meat) were pulverized by colloid mill, centrifuged to obtain a slurry, the solid content was adjusted to 25-30%, 0.1% sodium metabisulfite (to inhibit lipid oxidation) was added, and 0.2% (w / w) of neutral protease was added based on protein content. The temperature was adjusted to 42℃, the pH to 7, and the mixture was stirred at 100 rpm for 3 hours to prepare the protein.

[0042] (5) Mix the peanut cake fermented with Aspergillus niger with the fermented protein from the leftover food, add 0.2% of the total mass of the peanut cake fermented with Aspergillus niger and the fermented protein from the leftover food to the mixture, and hydrolyze at 45°C for 2 hours; then add phospholipid oil DHA and dynamic nutrient components, and homogenize at 10,000 rpm for 8 minutes to form an O / W emulsion; then add Antarctic krill powder, double-encapsulated astaxanthin microspheres, double-encapsulated Bacillus licheniformis and seaweed polysaccharide, mix well, freeze quickly at -80°C and then freeze-dry under vacuum, crush and sieve to obtain 200-500μm particles, which is the shrimp larvae feed.

[0043] Example 2

[0044] A shrimp larvae feed comprises the following components in parts by weight:

[0045] 25 portions of Aspergillus niger fermented peanut cake, 20 portions of meal waste fermented protein, 15 portions of Antarctic krill powder, 1.2 portions of double-encapsulated astaxanthin microspheres, 1.5 portions of double-encapsulated Bacillus licheniformis, 2 portions of seaweed polysaccharide, 3 portions of phospholipid oil DHA, and 6 portions of dynamic nutrient components;

[0046] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0047] The specific preparation method of the above-mentioned shrimp larvae feed is as follows:

[0048] (1) Preparation of peanut cake fermented with Aspergillus niger: The peanut cake was crushed to ≤2mm, the moisture content was adjusted to 50%, and it was steamed at 121℃ for 20min. Then, it was inoculated with Aspergillus niger (Aspergillus niger AS3.350, concentration 10). 6CFU / mL, inoculation amount of 1.5% v / w (commercially available), and 0.2% KH2PO4 were added. Solid fermentation was carried out at 30±1℃ for 40h, with stirring every 16h to prepare Aspergillus niger fermented peanut cake.

[0049] (2) Preparation of fermented protein from food waste: After sorting and crushing the food waste, inoculate it with a compound bacterial agent of Lactobacillus plantarum and Bacillus subtilis (Lactobacillus plantarum: Bacillus subtilis ≥ 3:1, with an addition amount of 5%), and ferment it in solid state at 39℃ for 48 hours to prepare fermented protein from food waste.

[0050] (3) Preparation of double-encapsulated astaxanthin microspheres and double-encapsulated Bacillus licheniformis: Mix astaxanthin or Bacillus licheniformis powder with 4% sodium alginate solution of astaxanthin or Bacillus licheniformis powder, add 2% CaCl2 solution of astaxanthin or Bacillus licheniformis powder, let stand for 15 min, solidify to form gel microspheres; immerse the gel microspheres in 1.5% chitosan acetate solution, adjust the pH to 5.8, stir for 35 min, take out the gel microspheres, drain, pre-freeze at -40℃ and vacuum dry to obtain double-encapsulated astaxanthin microspheres or double-encapsulated Bacillus licheniformis;

[0051] (4) Preparation of high cholesterol concentrate: Cuttlefish liver was enzymatically hydrolyzed with neutral protease (addition amount 0.5%) at 45℃ for 4 hours to release bound cholesterol and prepare cuttlefish liver enzymatic hydrolysate; the prepared cuttlefish liver enzymatic hydrolysate was mixed with shrimp shell powder (300 mesh) at a ratio of 1:2, and 5% by volume of phospholipid emulsifier (soy lecithin) was added to the mixture. The mixture was homogenized at 15000 rpm for 10 minutes to prepare high cholesterol concentrate;

[0052] Preparation of high-peptide protein: Fish processing by-products (fish head, viscera, and minced meat) were pulverized by colloid mill, centrifuged to obtain a slurry, the solid content was adjusted to 25-30%, 0.1% sodium metabisulfite (to inhibit lipid oxidation) was added, and 0.2% (w / w) of neutral protease was added based on protein content. The temperature was adjusted to 42℃, the pH to 7, and the mixture was stirred at 100 rpm for 3 hours to prepare the protein.

[0053] (5) Mix the peanut cake fermented with Aspergillus niger with the fermented protein from the leftover food, add 0.5% of the total mass of the peanut cake fermented with Aspergillus niger and the fermented protein from the leftover food to the mixture, and hydrolyze at 55°C for 2 hours; then add phospholipid oil DHA and dynamic nutrient components, and homogenize at 15000 rpm for 15 minutes to form an O / W emulsion; then add Antarctic krill powder, double-encapsulated astaxanthin microspheres, double-encapsulated Bacillus licheniformis and seaweed polysaccharide, mix well, freeze quickly at -80°C and then freeze-dry under vacuum, crush and sieve to obtain 200-500μm particles, which is the shrimp larvae feed.

[0054] Example 3

[0055] A shrimp larvae feed comprises the following components in parts by weight:

[0056] 22 portions of Aspergillus niger fermented peanut cake, 18 portions of meal waste fermented protein, 13 portions of Antarctic krill powder, 1.0 portion of double-encapsulated astaxanthin microspheres, 1.0 portion of double-encapsulated Bacillus licheniformis, 1.5 portions of seaweed polysaccharide, 2.5 portions of phospholipid oil DHA, and 4 portions of dynamic nutrient components.

[0057] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0058] The specific preparation method of the above-mentioned shrimp larvae feed is as follows:

[0059] (1) Preparation of peanut cake fermented with Aspergillus niger: The peanut cake was crushed to ≤2mm, the moisture content was adjusted to 49%, and it was steamed at 121℃ for 18min. Then, it was inoculated with Aspergillus niger (Aspergillus niger AS3.350, concentration 10). 6 CFU / mL, inoculation amount of 1.3% v / w (commercially available), and 0.2% KH2PO4 were added. Solid fermentation was carried out at 30±1℃ for 36h, with stirring every 14h to prepare Aspergillus niger fermented peanut cake.

[0060] (2) Preparation of fermented protein from food waste: After sorting and crushing the food waste, inoculate it with a compound bacterial agent of Lactobacillus plantarum and Bacillus subtilis (Lactobacillus plantarum: Bacillus subtilis ≥3:1, with an addition amount of 4%), and ferment it in solid state at 38℃ for 40h to prepare fermented protein from food waste.

[0061] (3) Preparation of double-embedded astaxanthin microspheres and double-embedded Bacillus licheniformis: Mix astaxanthin or Bacillus licheniformis powder with sodium alginate solution of 3.6% by weight of astaxanthin or Bacillus licheniformis powder, add 1.8% by weight of CaCl2 solution of astaxanthin or Bacillus licheniformis powder, let stand for 13 min, solidify to form gel microspheres; immerse the gel microspheres in 1.3% chitosan acetate solution, adjust the pH to 5.8, stir for 30 min, take out the gel microspheres, drain, pre-freeze at -40℃ and vacuum dry to obtain double-embedded astaxanthin microspheres or double-embedded Bacillus licheniformis;

[0062] (4) Preparation of high cholesterol concentrate: Cuttlefish liver was enzymatically hydrolyzed with neutral protease (addition amount 0.5%) at 45℃ for 4 hours to release bound cholesterol and prepare cuttlefish liver enzymatic hydrolysate; the prepared cuttlefish liver enzymatic hydrolysate was mixed with shrimp shell powder (300 mesh) at a ratio of 1:2, and 5% by volume of phospholipid emulsifier (soy lecithin) was added to the mixture. The mixture was homogenized at 15000 rpm for 10 minutes to prepare high cholesterol concentrate;

[0063] Preparation of high-peptide protein: Fish processing by-products (fish head, viscera, and minced meat) were pulverized by colloid mill, centrifuged to obtain a slurry, the solid content was adjusted to 25-30%, 0.1% sodium metabisulfite (to inhibit lipid oxidation) was added, and 0.2% (w / w) of neutral protease was added based on protein content. The temperature was adjusted to 42℃, the pH to 7, and the mixture was stirred at 100 rpm for 3 hours to prepare the protein.

[0064] (1) Mix peanut cake fermented with Aspergillus niger with leftover fermented protein, add 0.4% of papain of the total mass of peanut cake fermented with Aspergillus niger and leftover fermented protein, and hydrolyze at 50°C for 2 hours; then add phospholipid oil DHA and dynamic nutrient components, homogenize at 12000rpm for 10min to form an O / W emulsion; then add Antarctic krill powder, double-encapsulated astaxanthin microspheres, double-encapsulated Bacillus licheniformis and seaweed polysaccharide, mix well, freeze quickly at -80°C and then freeze dry under vacuum, crush and sieve to obtain 200-500μm particles, which is the shrimp larvae feed.

[0065] Comparative Example 1

[0066] A shrimp larvae feed comprises the following components in parts by weight:

[0067] 40 parts of Aspergillus niger fermented peanut cake, 13 parts of Antarctic krill powder, 1.0 part of double-encapsulated astaxanthin microspheres, 1.0 part of double-encapsulated Bacillus licheniformis, 1.5 parts of seaweed polysaccharide, 2.5 parts of phospholipid oil DHA, and 4 parts of dynamic nutrient components.

[0068] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0069] The preparation method is the same as in Example 3.

[0070] Comparative Example 2

[0071] A shrimp larvae feed comprises the following components in parts by weight:

[0072] The product contains 40 portions of fermented meal waste protein, 13 portions of Antarctic krill powder, 1.0 portion of double-encapsulated astaxanthin microspheres, 1.0 portion of double-encapsulated Bacillus licheniformis, 1.5 portions of seaweed polysaccharide, 2.5 portions of phospholipid oil DHA, and 4 portions of dynamic nutrient components.

[0073] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0074] The preparation method is the same as in Example 3.

[0075] Comparative Example 3

[0076] A shrimp larvae feed comprises the following components in parts by weight:

[0077] The product contains 13 parts Antarctic krill powder, 1.0 part double-encapsulated astaxanthin microspheres, 1.0 part double-encapsulated Bacillus licheniformis, 1.5 parts seaweed polysaccharide, 2.5 parts phospholipid oil DHA, and 4 parts dynamic nutrient components.

[0078] When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

[0079] The preparation method is the same as in Example 3.

[0080] Effect verification

[0081] To verify the technical effects of Examples 1-3 and Comparative Examples 1-3 of this application, healthy Litopenaeus vannamei larvae (initial weight 0.02±0.001g) were selected as experimental animals. The specific growth rate, feed conversion ratio, and survival rate of the Litopenaeus vannamei were measured during a 45-day culture period. The specific experimental procedures are as follows:

[0082] Experimental animals and grouping: Healthy Litopenaeus vannamei larvae (initial weight 0.02±0.001g) were selected and randomly divided into 5 groups (3 replicates per group, 100 shrimp per replicate).

[0083] Aquaculture system: recirculating aquaculture tank (300L), water temperature 30±0.5℃, salinity 25‰, continuous aeration;

[0084] Feeding method: Feed at 8% of the body weight of the animal, 4 times a day (8:00, 12:00, 16:00, 20:00), and spread evenly throughout the pond;

[0085] After the experiment, the specific growth rate, feed conversion ratio and survival rate of the above-mentioned whiteleg shrimp were measured respectively, and the results are shown in Table 1.

[0086] Table 1 Comparison of breeding effects in different groups

[0087] Group SGR(% / d) FCR Survival rate (%) Example 1 9.2±0.3 1.18±0.04 94.5±1.5 Example 2 9.4±0.3 1.15±0.05 95.2±1.3 Example 3 9.6±0.4 1.12±0.03 96.0±1.2 Comparative Example 1 8.0±0.2 1.48±0.08 84.3±2.0 Comparative Example 2 7.9±0.3 1.52±0.07 83.7±1.8 Comparative Example 3 6.5±0.4 1.85±0.10 72.1±2.5

[0088] Results Analysis: The specific growth rate, feed conversion ratio, and survival rate of Examples 1-3 of this application are all superior to those of Comparative Examples 1-3. Compared with Example 3, Comparative Example 1 replaced all FKR with FGNC, but its effect was not as good as that of Example 3. Compared with Example 3, Comparative Example 2 replaced all FGNC with FKR, but its effect was also not as good as that of Example 3. This shows that the combination of FGNC alone with other raw materials and the combination of FKR alone with other raw materials are not as good as the synergistic effect of FGNC and FKR with other raw materials, proving that there is a synergistic effect between the two.

[0089] Furthermore, the molting synchronicity, nitrogen utilization rate, and Vibrio challenge survival rate of Examples 1-3 and Comparative Examples 1-3 were determined (Vibrio challenge was performed using Vibrio parahaemolyticus ATCC17802 (10)). 5 (Soaking at CFU / mL for 30 minutes), the experimental results are shown in Table 2.

[0090] Table 2. Molting synchronicity, nitrogen utilization rate, and Vibrio challenge survival rate in different groups

[0091] Group Molting Synchronization CV% Nitrogen utilization rate (%) Vibrio survival rate after viral challenge (%) Example 1 15.2±0.8% 46.3±2.0 81.5±3.8 Example 2 12.0±0.6% 47.5±2.1 82.7±3.9 Example 3 13.5±0.7% 49.8±2.3 84.2±4.1 Comparative Example 1 32.5±1.5% 28.2±1.5 46.1±2.9 Comparative Example 2 34.8±1.8% 26.8±1.3 44.3±3.1 Comparative Example 3 48.3±2.2% 18.6±1.0 32.7±3.5

[0092] Results analysis: Compared with Examples 1-3, Comparative Example 1, using FGNC alone, lacked short-chain fatty acids (SCFAs), failed to activate the PepT1 transporter protein, and had a nitrogen utilization rate of only 28.2%.

[0093] Compared with Examples 1-3, Comparative Example 2 used FKR alone and lacked phytase, resulting in phytic acid residue, chelation of calcium / zinc, and asynchronous molting (CV% 34.8%).

[0094] To further verify the role of Aspergillus niger fermented peanut cake and meal waste fermented protein in fishmeal substitution, the traditional 100% fishmeal group was used as the control group. The fishmeal in the control group was replaced by Aspergillus niger fermented peanut cake (FGNC) and / or meal waste fermented protein (FKR) prepared in Example 3, as follows:

[0095] 100% Fishmeal Group: 60 parts steamed fishmeal, 8 parts Antarctic krill meal, 12 parts soybean meal, 10 parts flour, 4 parts fish oil, 0.3 parts astaxanthin (unencapsulated), 1.7 parts multivitamin premix, 2 parts mineral premix, and 2 parts squid paste;

[0096] 50% FGNC Substitute Group: Replace 30 portions of steamed fish meal with FGNC, and keep the rest unchanged;

[0097] 70% FGNC+FKR group: Replace 42 portions of steamed fish meal with FGNC+FKR (in the same proportion as the formulation in this application), and keep the rest unchanged.

[0098] The specific growth rate, feed conversion ratio, survival rate and nitrogen utilization rate of different groups were measured respectively. The experimental results are shown in Table 3.

[0099] Table 3. Results of measurements in different groups

[0100] Group 100% Fishmeal Group 50% FGNC Replacement Group 70% FGNC+FKR group SGR(% / d) 8.2±0.3 7.8±0.2 9.2±0.4 FCR 1.50 1.55 1.19 Survival rate (%) 82.3 80.1 93.6 Nitrogen utilization rate (%) 25.1 26.5 45.9

[0101] Results Analysis: Based on the aquatic feed industry standard (GB / T 22919-2008) and typical formulation design, this application designed a traditional fishmeal group formula. By adjusting the fishmeal replacement ratio, the application verified the role of Aspergillus niger fermented peanut cake and meal waste fermented protein in fishmeal replacement. Through the comparison of the above three groups, it can be seen that when 50% of FGNC alone is used to replace fishmeal, the effect after replacement is not as good as the 100% fishmeal group. This indicates that the 50% FGNC replacement has exceeded the replacement ratio, resulting in decreased shrimp growth and amino acid imbalance. However, when FGNC and FKR are combined, replacing fishmeal at a ratio of 70%, the effect is better than the 100% fishmeal group. This indicates that FGNC and FKR have a synergistic effect. The combination of the two can achieve a fishmeal replacement rate of 70% without growth inhibition. This breaks through the technical barrier of low plant protein replacement ratio of fishmeal in the existing technology, provides technical support for fishmeal replacement technology, and reduces production costs.

[0102] The various embodiments in this specification are described in a progressive manner, with each embodiment focusing on the differences from other embodiments. The same or similar parts between the various embodiments can be referred to each other.

[0103] The above description of the disclosed embodiments enables those skilled in the art to make or use the invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the invention. Therefore, the invention is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A shrimp larvae feed, characterized in that, It includes the following components in parts by weight: 20-25 parts fermented peanut cake with Aspergillus niger, 15-20 parts fermented protein from leftover food, 10-15 parts Antarctic krill powder, 0.5-1.2 parts double-encapsulated astaxanthin microspheres, 0.8-1.5 parts double-encapsulated Bacillus licheniformis, 1-2 parts seaweed polysaccharide, 2-3 parts phospholipid oil DHA, and 3-6 parts dynamic nutrient components.

2. The shrimp larvae feed according to claim 1, characterized in that, When the shrimp larvae are in the molting period, the dynamic nutrient composition is a high cholesterol concentrate; when the shrimp larvae are in the growth period, the dynamic nutrient composition is a high peptide protein.

3. The method for preparing shrimp larvae feed according to claim 1 or 2, characterized in that, Includes the following steps: (1) Preparation of peanut cake fermented by Aspergillus niger: crush peanut cake, adjust the moisture content to 48-50%, cook at 121℃ for 15-20 min under high pressure, inoculate with Aspergillus niger for solid-state fermentation, and prepare peanut cake fermented by Aspergillus niger. (2) Preparation of fermented protein from food waste: After sorting and crushing food waste, inoculate it with compound microbial agent and ferment it in solid state at 36-39℃ for 36-48h to prepare fermented protein from food waste. (3) Preparation of double-embedded astaxanthin microspheres and double-embedded Bacillus licheniformis: Mix astaxanthin or Bacillus licheniformis powder with sodium alginate solution, add CaCl2 solution dropwise, let stand for 10-15 min, and solidify to form gel microspheres; immerse the gel microspheres in 1.0-1.5% chitosan acetate solution, adjust the pH to 5.8, stir for 25-35 min, take out the gel microspheres, drain, pre-freeze at -40℃ and vacuum dry to obtain double-embedded astaxanthin microspheres or double-embedded Bacillus licheniformis; (4) Mix peanut cake fermented with Aspergillus niger with leftover fermented protein, add papain to it, and hydrolyze at 45-55℃ for 2 hours; then add phospholipid oil DHA and dynamic nutrient components, and homogenize at high speed to form an O / W emulsion; then add Antarctic krill powder, double-encapsulated astaxanthin microspheres, double-encapsulated Bacillus licheniformis and seaweed polysaccharide, mix well, freeze quickly at -80℃ and then freeze dry under vacuum, crush and screen to obtain 200-500μm particles, which is shrimp larvae feed.

4. The method for preparing shrimp larvae feed according to claim 3, characterized in that, In step (1), the peanut cake is crushed to ≤2mm; the concentration of Aspergillus niger is 106 CFU / mL, and the inoculum amount is 1-1.5% v / w; 0.2% KH2PO4 is added during the solid-state fermentation process; the solid-state fermentation temperature is 30±1℃, the fermentation time is 35-40h, and the stirring frequency is once every 12-16h.

5. The method for preparing shrimp larvae feed according to claim 3, characterized in that, The compound microbial agent mentioned in step (2) is a compound microbial agent of Lactobacillus plantarum and Bacillus subtilis, with a ratio of Lactobacillus plantarum to Bacillus subtilis ≥ 3:1; the amount of the compound microbial agent added is 3-5% of the amount of food waste.

6. The method for preparing shrimp larvae feed according to claim 3, characterized in that, In step (3), the amount of sodium alginate solution added is 3-4% of the mass of astaxanthin or Bacillus licheniformis powder; the amount of CaCl2 solution added is 1.5-2% of the mass of astaxanthin or Bacillus licheniformis powder.

7. The method for preparing shrimp larvae feed according to claim 3, characterized in that, The amount of papain added in step (4) is 0.2-0.5% of the total mass of peanut cake fermented with Aspergillus niger and fermented protein from leftover food; the speed of high-speed homogenization in step (4) is 10000-15000 rpm and the time is 8-15 min.

8. The method for preparing shrimp larvae feed according to claim 3, characterized in that, When the shrimp larvae are in the molting period, the dynamic nutrient component is a high-cholesterol concentrate, which is prepared by emulsifying shrimp shell powder and enzymatically hydrolyzed squid liver solids at a weight ratio of 2:

1. When the shrimp larvae are in the growth period, the dynamic nutrient component is a high-peptide protein, which is prepared by enzymatically hydrolyzing fish lysate at 42-46℃ for 3-5 hours.

9. The application of the shrimp larvae feed according to claim 1 or 2 or the shrimp larvae feed prepared by any of the preparation methods according to claims 3-8 in the rearing process of Litopenaeus vannamei.

10. The combination of Aspergillus niger fermented peanut cake and meal waste fermented protein prepared in steps (1) and (2) of claim 3, used as a fishmeal substitute in the preparation of shrimp larvae feed.