Mikania micrantha dry powder as well as preparation method and application thereof, eel feed as well as preparation method and application thereof

Abstract

The invention provides mikania micrantha dry powder as well as a preparation method and application thereof, eel feed as well as a preparation method and application thereof, and relates to the technical field of aquaculture. Specifically, to-be-fermented mikania micrantha powder is prepared from fresh mikania micrantha stalks and leaves, mikania micrantha zymophyte powder is prepared from fresh mikania micrantha leaves and cotton aphid adults, then mikania micrantha zymophyte liquid and a mikania micrantha liquid fermentation culture medium with specific components are prepared respectively, and the novel mikania micrantha dry powder is obtained through fermentation culture. In addition, the novel eel feed is prepared from mikania micrantha dry powder, white fish meal, alpha-starch, choline chloride, calcium hydrophosphate, fish oil, a compound additive, compound vitamins, compound minerals and an adhesive. The problems of eel fry breeding, artificial breeding and bait supply can be solved, and the artificial eel breeding cost is greatly reduced.

Description

Technical Field

[0001] The present invention relates to the technical field of aquaculture, and in particular to micrantha dry powder, a preparation method and application thereof, and eel feed, a preparation method and application thereof. Background Art

[0002] Mikania micrantha is a perennial herb or shrubby climbing vine of the genus Eupatorium in the Asteraceae family, native to South and Central America. It has become one of the most serious invasive alien species in tropical and subtropical regions worldwide. Current progress in artificial control, chemical control, and biological control has failed to fundamentally curb the spread and spread of Mikania micrantha. Therefore, the most effective way to control the natural spread of Mikania micrantha is to find ways to effectively utilize it. Mikania micrantha is high in crude fiber and contains a certain amount of protein, vitamins, and minerals. As a component of compound feed, it can meet the growth and development needs of farmed animals. Currently, research on the application of Mikania micrantha focuses on control techniques, allelopathic effects, and insect breeding. There are few reports on the development and utilization of Mikania micrantha in aquatic animal feed, especially in carnivorous fish feed.

[0003] Eels are carnivorous fish with a high protein requirement. my country accounts for approximately 80% of the global eel aquaculture sector, and most commercial eel farming relies on imported white fish meal as a key protein feed, with a high usage rate. Currently, there have been no trials of low-protein compound feeds that replace protein feed with energy feed in artificial eel farming. The "First Batch of National Aquatic Breeding Joint Public Relations on Varieties and Key Tasks" document states that the cultivation of reserve broodstock and newly hatched larvae are key to the mass production of artificially propagated lancelet eels. However, experimental research by the applicant has revealed that, under the same farming conditions, the economic cost of feeding eel broodstock a compound feed whose primary protein feed is fish meal is significantly higher than feeding a compound feed that partially replaces fish meal with energy feed. Furthermore, regardless of the commercial feed available on the market, the economic cost of artificial eel breeding remains high. This is primarily due to the high deformity rate of newly hatched larvae at the lancelet eel stage, making it difficult for them to mature into the glass eel stage. Therefore, in order to reduce the cost of artificial eel feed and artificial breeding, and to achieve the goal of reducing the cost of commercial eel breeding, finding high-efficiency energy feed raw materials that can replace part of fish meal is the key to solving the problem.

[0004] Existing technology indicates that adding Mikania micrantha directly to feed can cause the animals to die from ingesting toxic substances such as sesquiterpene lactone compounds naturally present in Mikania micrantha. Therefore, if Mikania micrantha is to be used as an economical energy feed ingredient for eels and to be widely adopted, it must at least have a simple process, low cost, stable effects, and low toxicity or detoxification properties.

[0005] In view of this, the present invention is proposed. Summary of the Invention

[0006] The first object of the present invention is to provide a method for preparing dry powder of Mikania micrantha, which is used to reduce the content of toxic substances naturally contained in Mikania micrantha, so as to overcome the defect that Mikania micrantha or its extracts are toxic to the body of the cultivated subjects.

[0007] The second object of the present invention is to provide a dry powder of Mikania micrantha.

[0008] The third purpose of the present invention is to provide an eel feed, which, on the one hand, solves the problem of high artificial breeding costs at the current stage from the perspective of reducing the deformity rate of newly hatched artificially bred eels during the eel stage; on the other hand, it seeks the possibility of low-protein eel feed, from the perspective of replacing protein feed with energy feed, to solve the problem of high feed costs of artificial feed breeding eels at the current stage.

[0009] A fourth object of the present invention is to provide a method for preparing eel feed.

[0010] The fifth object of the present invention is to provide use of the micrantha dry powder or the eel feed in aquaculture.

[0011] In order to achieve the above-mentioned purpose of the present invention, the following technical solutions are adopted:

[0012] A method for preparing micrantha dry powder comprises the following steps:

[0013] (1) preparing fresh Mikania micrantha stems and fresh Mikania micrantha leaves, drying, crushing, and sieving to obtain Mikania micrantha stem and leaf powder; then fully mixing the Mikania micrantha stem and leaf powder, cellulase, and sodium bicarbonate solution, and performing a first reaction to separate the solid and liquid to obtain Mikania micrantha hydrolyzate; fully mixing the Mikania micrantha hydrolyzate, sodium hydroxide solution, and sodium sulfite solution, and performing a second reaction to adjust the pH to 6.9-7.1, and then performing reduced pressure evaporation, cooling, and removing surface crystals to obtain Mikania micrantha thick paste; fully mixing the Mikania micrantha thick paste, sodium alginate, and urea, and performing freeze-drying, crushing, and sieving to obtain Mikania micrantha fermentation powder;

[0014] (2) placing adult cotton aphids on fresh Mikania micrantha leaves and aseptically cultivating them for 2 to 5 days, then crushing the insect bodies to obtain Mikania micrantha aphid paste; mixing the Mikania micrantha aphid paste, anhydrous ethanol, n-hexane, and gluten for fermentation, and after the fermentation is completed, standing, drying in the dark, crushing, and sieving to obtain Mikania micrantha fermentation powder;

[0015] (3) preparing the micrantha fermentation bacteria powder, bream larvae powder, calcium hydrogen phosphate, potassium aluminum sulfate dodecahydrate, disodium hydrogen phosphate solution and sodium dihydrogen phosphate solution, and fully mixing to obtain the micrantha fermentation bacteria liquid;

[0016] (4) preparing an aqueous solution of ferrous chloride, magnesium sulfate, manganese chloride, γ-aminobutyric acid, sodium tetraborate decahydrate, and lotus root starch as a liquid fermentation medium for Mikania micrantha;

[0017] (5) The micrantha to-be-fermented powder and the micrantha fermentation broth are uniformly mixed, inoculated into the micrantha liquid fermentation medium and fermented, and the micrantha fermentation broth, corn flour, alfalfa powder, kudzu powder, and fermented palm meal are regularly added; after the fermentation is completed, the tank is placed, ammonium chloride is added to the fermentation product and the mixture is fully mixed, and then a solid phase is obtained by solid-liquid separation, and the moisture of the solid phase is removed to obtain the micrantha dry powder.

[0018] Preferably, step (1) includes at least one of the following features (a) to (f):

[0019] (a) the mass ratio of the fresh Mikania micrantha stems to the fresh Mikania micrantha leaves is (30-60):(10-16);

[0020] (b) the mass ratio of the Mikania micrantha stem and leaf powder, the cellulase and the sodium bicarbonate solution is (84-115): (3-5): (525-702);

[0021] (c) the mass ratio of the Mikania micrantha enzymatic hydrolysate, the sodium hydroxide solution and the sodium sulfite solution is (286-502): (33-55): (1.7-3.9);

[0022] (d) the mass ratio of the micrantha micrantha slurry, the sodium alginate and the urea is (90-120):(21-27):(0.14-2.22);

[0023] (e) The conditions of the first reaction include: a temperature of 28° C. to 43° C. and a duration of 48 h to 72 h;

[0024] (f) The conditions of the second reaction include: temperature of 110° C. to 130° C., and duration of 1.8 h to 3 h.

[0025] Preferably, step (2) includes at least one of the following features (a) to (c):

[0026] (a) The sterile culture conditions include: a temperature of 18°C ​​to 24°C, a light intensity of 10,000 lux to 20,000 lux, and a light duration L:D of (10 to 14) h:(10 to 14) h;

[0027] (b) the mass ratio of the Mikania micrantha aphid paste, the anhydrous ethanol, the n-hexane and the gluten is (160-228): (34-72): (1-6): (13-28);

[0028] (c) The fermentation conditions include: a temperature of 22° C. to 30° C., a fermentation time of 48 to 60 hours, and exhaust gas for 5 to 10 minutes every 3 hours after the 18th hour of the fermentation.

[0029] Preferably, in step (3), the fermented bacteria liquid of Mikania micrantha comprises the following components in parts by weight: 11 to 23 parts of the fermented bacteria powder of Mikania micrantha, 4 to 8 parts of the bream fry powder, 15 to 19 parts of the calcium hydrogen phosphate, 5 to 12 parts of the potassium aluminum sulfate dodecahydrate, 89 to 104 parts of the disodium hydrogen phosphate solution, and 73 to 95 parts of the sodium dihydrogen phosphate solution.

[0030] Preferably, in step (4), the aqueous solution of the liquid fermentation medium of Mikania micrantha comprises the following components in parts by weight: 17 to 33 parts of ferrous chloride, 37.5 to 45.5 parts of magnesium sulfate, 14.7 to 24.3 parts of manganese chloride, 0.3 to 0.7 parts of γ-aminobutyric acid, 21 to 30 parts of sodium tetraborate decahydrate, 174 to 191 parts of lotus root starch, and 1000 to 1200 parts of water.

[0031] Preferably, in step (5), 128 to 360 parts of the Mikania micrantha fermentation powder and 100 to 110 parts of the Mikania micrantha fermentation bacterial liquid are mixed evenly by mass, and inoculated into 818 to 1205 parts of the Mikania micrantha liquid fermentation medium; after the fermentation is completed, the tank is placed, and 16 to 19 parts of ammonium chloride are added to the fermentation product and mixed thoroughly.

[0032] Preferably, the fermentation culture conditions include: a temperature of 14° C. to 20° C., a duration of 7 to 16 days, and exhaust every 6 to 8 hours.

[0033] A micrantha dry powder is prepared based on the preparation method of the micrantha dry powder.

[0034] An eel feed comprising the following ingredients in parts by weight:

[0035] The present invention comprises 41-47 parts of micrantha dry powder, 28-34 parts of white fish meal, 20-25 parts of α-starch, 0.1-0.3 parts of choline chloride, 0.1-0.3 parts of calcium hydrogen phosphate, 0.2-0.5 parts of fish oil, 0.2-0.4 parts of composite additives, 0.2-0.4 parts of composite vitamins, 0.2-0.4 parts of composite minerals and 0.1-0.3 parts of adhesive.

[0036] Preferably, it includes at least one of the following features (A) to (C):

[0037] (A) The composite additive comprises the following components by mass: 12 to 15 parts of β-glucanase, 9 to 17 parts of β-xylosidase, 35 to 56 parts of β-D-glucose oxidoreductase, 5 to 26 parts of α-galactosidase, 6 to 16 parts of bream fry meal, and 14 to 28 parts of the fermented bacteria powder of Mikania micrantha;

[0038] (B) the vitamin complex comprises the following ingredients, calculated by mass: 22-30 parts of vitamin A, 40-65 parts of vitamin C, 0.2-0.45 parts of riboflavin, 0.07-0.18 parts of folic acid, 0.03-0.06 parts of vitamin B1, 0.3-0.9 parts of biotin, 125-238 parts of vitamin D3, 12-17 parts of thiamine, 32-4 parts of vitamin K, 7-11 parts of vitamin E, 22-28 parts of vitamin B6, 80-100 parts of inositol, 15-29 parts of pantothenic acid, and 63-74 parts of niacinamide;

[0039] (C) The composite mineral comprises the following components in parts by mass: 15-19 parts of zinc chloride, 4-15 parts of calcium iodate, 0.2-0.5 parts of potassium fluoride, 12-14 parts of copper sulfate, 0.14-0.18 parts of sodium molybdate, 137-142 parts of manganese chloride, 172-190 parts of magnesium sulfate, 1-1.2 parts of sodium selenite, 11.3-12 parts of cobalt sulfate and 330-365 parts of ferrous sulfate.

[0040] A method for preparing eel feed comprises the following steps:

[0041] The solid phase components of the eel feed are crushed and sieved respectively, and then the components of the eel feed are fully mixed and granulated to obtain the eel feed.

[0042] Use of the micrantha dry powder and the eel feed in aquaculture.

[0043] Compared with the prior art, the present invention has the following beneficial effects:

[0044] (1) The present invention provides a dry powder of Mikania micrantha mainly used for aquatic feed, which is obtained by using adult cotton aphids that have digested Mikania micrantha leaves and containing chyme as the bacterial source, and using Mikania micrantha to be fermented powder as the main raw material, supplemented with triangular bream fry meal, corn meal, alfalfa meal, kudzu powder, and fermented palm meal. The powder is fermented in a Mikania liquid fermentation medium with specific components, expanded and cultured, and then dried. The present invention simulates the process of decomposing and transforming toxic substances naturally contained in Mikania micrantha in the body of adult cotton aphids to the greatest extent possible, while fully converting the crude fiber rich in Mikania micrantha into energy feed before being added to the feed components, and preliminarily decomposing proteins and other non-starch polysaccharides to form amino acids and oligosaccharides, which are more conducive to animal digestion and absorption, and improve the animal weight gain rate.

[0045] (2) The present invention provides an eel feed, which includes a specific composite additive, and the composite additive includes fermented fungus powder of Mikania micrantha; the present invention is the first to completely add the fungus powder derived from the Aphids flora to the feed additive of carnivorous fish, and explore a new addition ratio of feed enzymes within a new enzyme activity range suitable for eels; the eel feed of the present invention can meet the olfactory and taste requirements of eels during the process of acclimating them to feed, greatly reduce the deformity rate of artificially bred offspring of eels in the willow leaf eel stage, and improve the feeding rate of eels.

[0046] (3) The eel feed provided by the present invention has a reasonable formula, a wide range of raw materials, and is inexpensive. Most of the raw materials are essential nutrients for eel growth. While reducing the cost of feed raw materials, they also fully utilize their nutritional functions, can promote eel growth, and improve feed efficiency. At the same time, the preparation method of the eel feed of the present invention is simple and easy, with low preparation cost, suitable for mass production, and has significant social and economic benefits. DETAILED DESCRIPTION

[0047] The technical solution of the present invention will be clearly and completely described below in conjunction with specific embodiments, but those skilled in the art will understand that the embodiments described below are part of embodiments of the present invention, rather than all embodiments, and are only used to illustrate the present invention, and should not be regarded as limiting the scope of the present invention. Based on the embodiments in the present invention, all other embodiments obtained by those of ordinary skill in the art without making creative work are within the scope of protection of the present invention. If specific conditions are not specified in the embodiments, they are carried out according to conventional conditions or the conditions recommended by the manufacturer. If the manufacturer is not specified for the reagents or instruments used, they are all conventional products that can be purchased commercially. In addition, the terms "first", "second" and "third" are only used for descriptive purposes and are not to be understood as indicating or implying relative importance.

[0048] The first aspect of the present invention is to provide a method for preparing micrantha dry powder, which mainly comprises the following steps (1) to (5).

[0049] It should be noted that any "full mixing" or "reaction" described in the present invention can be assisted by oscillation, stirring, shaking, centrifugation, ultrasound, heating, etc.; any "solid-liquid separation" described in the present invention can be carried out by decantation, filtration, centrifugation, filter or membrane separation.

[0050] (1) preparing fresh Mikania micrantha stems and fresh Mikania micrantha leaves, drying, crushing and sieving to obtain Mikania micrantha stem and leaf powder; then fully mixing the Mikania micrantha stem and leaf powder, cellulase and sodium bicarbonate solution, and performing a first reaction to separate the solid and liquid to obtain Mikania micrantha hydrolysate; fully mixing the Mikania micrantha hydrolysate, sodium hydroxide solution and sodium sulfite solution, and performing a second reaction to adjust the pH to 6.9-7.1, and then performing reduced pressure evaporation, cooling and removing surface crystals to obtain Mikania micrantha thick paste; fully mixing the Mikania micrantha thick paste, sodium alginate and urea, and performing freeze-drying, crushing and sieving to obtain Mikania micrantha fermentation powder.

[0051] As a preferred embodiment, the mass ratio of the fresh Mikania micrantha stems to the fresh Mikania micrantha leaves is (30-60):(10-16).

[0052] As a preferred embodiment, the drying of the fresh Mikania micrantha stems and the fresh Mikania micrantha leaves is carried out under light-proof conditions. In a more ...

[0053] As a preferred embodiment, the Mikania micrantha stem and leaf powder is obtained by sieving through a 160-200 mesh sieve; or, the particle size of the Mikania micrantha stem and leaf powder is 70 μm to 120 μm.

[0054] As a preferred embodiment, the mass ratio of the Mikania micrantha stem and leaf powder, the cellulase and the sodium bicarbonate solution is (84-115): (3-5): (525-702).

[0055] As a preferred embodiment, the enzymatic activity of the cellulase is 12000U to 21500U.

[0056] As a preferred embodiment, the concentration of the sodium bicarbonate solution is 0.04 mol / L to 0.08 mol / L; in some more preferred embodiments, the pH of the sodium bicarbonate solution is 8 to 8.2.

[0057] As a preferred embodiment, the conditions of the first reaction include: temperature of 28°C to 43°C, and duration of 48h to 72h.

[0058] As a preferred embodiment, the mass ratio of the Mikania micrantha enzymatic hydrolyzate, the sodium hydroxide solution and the sodium sulfite solution is (286-502): (33-55): (1.7-3.9).

[0059] As a preferred embodiment, the concentration of the sodium hydroxide solution is 0.2 mol / L to 0.4 mol / L.

[0060] As a preferred embodiment, the concentration of the sodium sulfite solution is 0.75 mol / L to 0.9 mol / L.

[0061] As a preferred embodiment, the conditions of the second reaction include: temperature of 110° C. to 130° C., and duration of 1.8 h to 3 h.

[0062] As a preferred embodiment, the pH is adjusted to 6.95-7.05.

[0063] As a preferred embodiment, the conditions for the reduced pressure evaporation include: a temperature of 65° C. to 80° C., and a duration of 5 min to 7 min.

[0064] As a preferred embodiment, the cooling temperature is 4°C to 10°C.

[0065] As a preferred embodiment, the mass ratio of the micrantha micrantha slurry, the sodium alginate and the urea is (90-120): (21-27): (0.14-2.22).

[0066] As a preferred embodiment, the freeze-drying temperature is -120°C to -80°C.

[0067] As a preferred embodiment, the Mikania micrantha to be fermented powder is obtained by sieving through a 50-80 mesh sieve; or, the particle size of the Mikania micrantha to be fermented powder is 170 μm to 290 μm.

[0068] (2) placing adult cotton aphids on fresh Mikania micrantha leaves and carrying out sterile culture for 2 to 5 days, taking out the insect bodies and crushing them to obtain Mikania micrantha aphid paste; mixing the Mikania micrantha aphid paste, anhydrous ethanol, n-hexane and gluten for fermentation, and after the fermentation is completed, standing, drying in the dark, crushing and sieving to obtain Mikania micrantha fermentation powder.

[0069] As a preferred embodiment, the cotton aphid adults are selected from a sterile breeding type, and are kept in an empty stomach before being placed.

[0070] As a preferred embodiment, the placement density of the cotton aphid adults is: 2 ~5cm 2 1 to 2 cotton aphid adults are placed on the leaves.

[0071] As a preferred embodiment, after the placement of the cotton aphid adults, a colorless transparent bag with holes is placed on the fresh Mikania micrantha leaves.

[0072] As a preferred embodiment, the conditions for the aseptic culture include: a temperature of 18°C ​​to 24°C, a light intensity of 10,000 lux to 20,000 lux, and a light duration L:D (i.e., the ratio of light time to dark time) of (10 to 14) h: (10 to 14) h.

[0073] As a preferred embodiment, the mass ratio of the Mikania micrantha aphid paste, the anhydrous ethanol, the n-hexane and the gluten is (160-228): (34-72): (1-6): (13-28).

[0074] As a preferred embodiment, the fermentation conditions include: a temperature of 22° C. to 30° C., a fermentation time of 48 h to 60 h; and exhaust gas is performed for 5 min to 10 min every 3 h after the 18th hour of fermentation.

[0075] As a preferred embodiment, the standing conditions include: a temperature of 20°C to 30°C (room temperature) and a duration of 6h to 8h.

[0076] As a preferred embodiment, the temperature for drying in the dark is 8°C to 15°C.

[0077] As a preferred embodiment, the fermented bacteria powder of Mikania micrantha is obtained by sieving through a 60-100 mesh sieve; or, the particle size of the fermented bacteria powder of Mikania micrantha is 140 μm to 250 μm.

[0078] (3) preparing the Mikania micrantha fermentation bacteria powder, bream fry powder, calcium hydrogen phosphate, potassium aluminum sulfate dodecahydrate, disodium hydrogen phosphate solution and sodium dihydrogen phosphate solution, and fully mixing them to obtain Mikania micrantha fermentation bacteria liquid.

[0079] As a preferred embodiment, the preparation method of the bream fry meal comprises: crushing newly hatched bream fry into a minced state, and drying, crushing, and sieving to obtain the bream fry meal.

[0080] As a preferred embodiment, the bream fry meal is obtained by sieving through a 60-100 mesh sieve to 90%; or, the particle size of the Mikania micrantha fermented bacteria powder is 140 μm to 250 μm.

[0081] As a preferred embodiment, the fermented bacteria liquid of Mikania micrantha includes the following components in parts by weight: 11 to 23 parts of the fermented bacteria powder of Mikania micrantha, 4 to 8 parts of the bream fry powder, 15 to 19 parts of the calcium hydrogen phosphate, 5 to 12 parts of the potassium aluminum sulfate dodecahydrate, 89 to 104 parts of the disodium hydrogen phosphate solution, and 73 to 95 parts of the sodium dihydrogen phosphate solution.

[0082] As a preferred embodiment, the concentration of the disodium hydrogen phosphate solution is 0.05 mol / L to 0.09 mol / L.

[0083] As a preferred embodiment, the concentration of the sodium dihydrogen phosphate solution is 0.7 mol / L to 0.9 mol / L.

[0084] (4) An aqueous solution of ferrous chloride, magnesium sulfate, manganese chloride, γ-aminobutyric acid, sodium tetraborate decahydrate and lotus root starch was prepared as a liquid fermentation medium for Mikania micrantha.

[0085] As a preferred embodiment, the aqueous solution of the liquid fermentation medium of Mikania micrantha includes the following components in parts by weight: 17 to 33 parts of ferrous chloride, 37.5 to 45.5 parts of magnesium sulfate, 14.7 to 24.3 parts of manganese chloride, 0.3 to 0.7 parts of γ-aminobutyric acid, 21 to 30 parts of sodium tetraborate decahydrate, 174 to 191 parts of lotus root starch, and 1000 to 1200 parts of water.

[0086] (5) The micrantha to-be-fermented powder and the micrantha fermentation broth are uniformly mixed, inoculated into the micrantha liquid fermentation medium and fermented, and the micrantha fermentation broth, corn flour, alfalfa powder, kudzu powder, and fermented palm meal are regularly added; after the fermentation is completed, the tank is placed, ammonium chloride is added to the fermentation product and the mixture is fully mixed, and then a solid phase is obtained by solid-liquid separation, and the moisture of the solid phase is removed to obtain the micrantha dry powder.

[0087] As a preferred embodiment, 128-360 parts of the Mikania micrantha fermentation powder and 100-110 parts of the Mikania micrantha fermentation liquid are uniformly mixed by mass and inoculated into 818-1205 parts of the Mikania micrantha liquid fermentation medium.

[0088] As a preferred embodiment, the fermentation culture conditions include: a temperature of 14° C. to 20° C., a duration of 7 to 16 days; and exhaust every 6 to 8 hours.

[0089] As a more preferred embodiment, the conditions for regularly adding materials during the fermentation culture include: 1) adding 30-50 parts of the Mikania micrantha fermentation broth and 10-16 parts of the corn flour daily; 2) adding 26-32 parts of alfalfa meal in four batches at 6-hour intervals on the fourth day of fermentation (i.e., adding 1 / 4 of the total amount each time); 3) adding 7-9 parts of kudzu root every two days; and 4) adding 50-55 parts of fermented palm meal every three days. It is worth noting that the mass parts in this embodiment are calculated based on 128-360 parts of the Mikania micrantha fermentation powder, 100-110 parts of the Mikania micrantha fermentation broth, and 818-1205 parts of the Mikania micrantha liquid fermentation medium.

[0090] As a preferred embodiment, the fermented palm meal is obtained by screening through a 60-mesh to 100-mesh sieve; or, the particle size of the fermented palm meal is 140 μm to 250 μm.

[0091] As a preferred embodiment, based on 128-360 parts of the Mikania micrantha fermentation powder, 100-110 parts of the Mikania micrantha fermentation bacteria liquid, and 818-1205 parts of the Mikania micrantha liquid fermentation medium, the added amount of ammonium chloride is 16-19 parts.

[0092] The second aspect of the present invention is to provide Mikania micrantha dry powder prepared by the preparation method of Mikania micrantha dry powder as described in the first aspect.

[0093] The third aspect of the present invention is to provide an eel feed, comprising the dry powder of Mikania micrantha as described in the second aspect.

[0094] Specifically, the eel feed includes the following ingredients in parts by weight: 41 to 47 parts of micrantha dry powder, 28 to 34 parts of white fish meal, 20 to 25 parts of α-starch, 0.1 to 0.3 parts of choline chloride, 0.1 to 0.3 parts of calcium hydrogen phosphate, 0.2 to 0.5 parts of fish oil, 0.2 to 0.4 parts of composite additives, 0.2 to 0.4 parts of composite vitamins, 0.2 to 0.4 parts of composite minerals, and 0.1 to 0.3 parts of adhesives.

[0095] As some optional embodiments, the weight proportions of the components in the eel feed include but are not limited to: 41, 42, 43, 44, 45, 46, 47 of the micrantha dry powder; 28, 29, 30, 31, 32, 33, 34 of white fish meal; 20, 21, 22, 23, 24, 25 of α-starch; 0.1, 0.15, 0.2, 0.25, 0.3 of choline chloride; 0.1, 0.15, 0.2, 0.25, 0.3 of calcium hydrogen phosphate; 0.2, 0.25, 0.3, 0.3 of fish oil. 35, 0.4, 0.45, 0.5; compound additives 0.2, 0.25, 0.3, 0.35, 0.4; compound vitamins 0.2, 0.25, 0.3, 0.35, 0.4; compound minerals 0.2, 0.25, 0.3, 0.35, 0.4; adhesives 0.1, 0.15, 0.2, 0.25, 0.3; all units are in parts, and the weight parts of each component in the eel feed can be selected from the point values ​​listed above, or the numerical range formed by any two point values.

[0096] As a preferred embodiment, the composite additive includes the following components in parts by mass: 12 to 15 parts of β-glucanase, 9 to 17 parts of β-xylosidase, 35 to 56 parts of β-D-glucose oxidoreductase, 5 to 26 parts of α-galactosidase, 6 to 16 parts of triangular bream fry meal and 14 to 28 parts of the fermented bacteria powder of micrantha.

[0097] As a more preferred embodiment, the enzymatic activity of the β-glucanase is 78,000 U / g to 130,000 U / g.

[0098] As a more preferred embodiment, the enzyme activity of the β-xylosidase is 228,000 U / g to 360,000 U / g.

[0099] As a more preferred embodiment, the enzyme activity of the β-D-glucose oxidoreductase is 6730 U / g to 10800 U / g.

[0100] As a more preferred embodiment, the enzyme activity of the α-galactosidase is 7500 U / g to 9600 U / g.

[0101] As a more preferred embodiment, the preparation method of the composite additive includes: taking the α-galactosidase, adding the triangular bream fry powder, the β-xylosidase, the β-glucanase, the micrantha fermentation powder, and the β-D-glucose oxidoreductase in sequence, mixing each raw material after adding, and fully mixing after the addition is completed.

[0102] As a preferred embodiment, the multivitamin includes the following ingredients in parts by mass: 22-30 parts of vitamin A, 40-65 parts of vitamin C, 0.2-0.45 parts of riboflavin, 0.07-0.18 parts of folic acid, 0.03-0.06 parts of vitamin B1, 0.3-0.9 parts of biotin, 125-238 parts of vitamin D3, 12-17 parts of thiamine, 32-4 parts of vitamin K, 7-11 parts of vitamin E, 22-28 parts of vitamin B6, 80-100 parts of inositol, 15-29 parts of pantothenic acid and 63-74 parts of niacinamide.

[0103] As a more preferred embodiment, the preparation method of the multivitamin includes: taking vitamin B12, and adding folic acid, riboflavin, biotin, vitamin K3, vitamin E, thiamine, pantothenic acid, vitamin B6, vitamin A, vitamin C, niacinamide, inositol, and vitamin D3 in sequence, mixing each raw material after adding, and fully mixing after the addition is completed.

[0104] As a preferred embodiment, the composite mineral includes the following components in parts by mass: 15-19 parts of zinc chloride, 4-15 parts of calcium iodate, 0.2-0.5 parts of potassium fluoride, 12-14 parts of copper sulfate, 0.14-0.18 parts of sodium molybdate, 137-142 parts of manganese chloride, 172-190 parts of magnesium sulfate, 1-1.2 parts of sodium selenite, 11.3-12 parts of cobalt sulfate and 330-365 parts of ferrous sulfate.

[0105] As a more preferred embodiment, the preparation method of the composite mineral comprises: taking the sodium molybdate, and sequentially adding the potassium fluoride, the sodium selenite, the calcium iodate, the cobalt sulfate, the copper sulfate, the zinc chloride, the manganese chloride, the magnesium sulfate, and the ferrous sulfate, mixing each raw material after adding, and fully mixing after the addition is completed.

[0106] As a preferred embodiment, the adhesive includes kelp glue, diatomaceous earth or xanthan gum.

[0107] The fourth aspect of the present invention is to provide a method for preparing the eel feed as described in the third aspect, which mainly comprises the following steps: crushing and sieving the solid phase components of the eel feed separately, fully mixing the various components of the eel feed, and then granulating to obtain the eel feed.

[0108] As a preferred embodiment, the solid phase components of the eel feed are sieved through a 100-140 mesh sieve.

[0109] As a preferred embodiment, each raw material is mixed after being added, and the mixture is fully mixed after the addition is completed.

[0110] As a preferred embodiment, the eel feed is in the form of columnar particles, the diameter of the eel feed is 1 / 10 to 1 / 7 of the eel's mouth, and the ratio of diameter to height is (1 to 3): (6 to 9).

[0111] As a preferred embodiment, the eel feed also includes some water, with a water content of 20% to 35%.

[0112] The fifth aspect of the present invention is to provide uses of the dry powder of Mikania micrantha described in the second aspect and the eel feed described in the third aspect in aquaculture.

[0113] Example 1

[0114] (1) By weight, 30 parts of fresh, unrotten Mikania micrantha stems and 10 parts of fresh, unrotten Mikania micrantha leaves were taken, dried in the dark at 16° C., and crushed until 100% passed through a 180-mesh sieve to obtain Mikania micrantha stem and leaf powder.

[0115] (2) By weight, 84 parts of Mikania micrantha stem and leaf powder, 3 parts of cellulase with an enzyme activity of 12,000 U, and 525 parts of 0.05 mol / L sodium bicarbonate solution with a pH of 8 were mixed. The mixture was shaken at 28°C and 60 rpm for 48 hours. The mixture was then centrifuged at 4°C and 4,000 rpm for 5 minutes, and the supernatant was collected to obtain Mikania micrantha enzymatic hydrolyzate.

[0116] (3) Mix 286 parts of Mikania micrantha enzymatic hydrolysate, 33 parts of 0.2 mol / L sodium hydroxide solution, and 1.7 parts of 0.75 mol / L sodium sulfite solution by weight. Treat at 110°C and 120 r / min for 1.8 h, then cool to room temperature. After adjusting the pH to 6.95, evaporate under reduced pressure at 65°C for 5 min, then cool to 4°C and remove surface crystals to obtain a Mikania micrantha slurry.

[0117] (4) Take 90 parts by weight of Mikania micrantha thick paste, 21 parts of sodium alginate, and 0.14 parts of urea, mix them evenly, freeze-dry at -120°C, and grind them at low temperature until 100% of the powder passes through a 60-mesh sieve to obtain Mikania micrantha powder to be fermented.

[0118] (5) On the fresh, unrotten leaves of healthy Mikania plants, press 1 head: 5cm 2 Place sterile, empty-bellied cotton aphids at a density of 10000 lux and place them in colorless, transparent bags with holes. Then, culture them for two days at 18°C, 10,000 lux, and a photoperiod of 10:14 hours. On the final day, remove the bags, remove all insects, and mash them into a paste to obtain the micrantha aphid paste.

[0119] (6) 160 parts by weight of Mikania micrantha aphid paste, 34 parts by weight of anhydrous ethanol, 1 part by weight of n-hexane, and 13 parts by weight of gluten were placed in a fermentation tank. After fermentation at 22°C for 18 hours, the mixture was vented every 3 hours for 5 minutes. The fermentation tank was then opened after 48 hours. The fermented mixture was allowed to stand at room temperature for 6 hours, then dried in the dark at 8°C. The mixture was then ground into powder until 100% of the powder passed through an 80-mesh sieve to obtain Mikania micrantha fermentation powder.

[0120] (7) Take newly hatched bream fry, crush them into a paste, dry them at 100° C., grind them into powder at room temperature until 90% of the powder passes through an 80-mesh sieve, and obtain bream fry powder.

[0121] (8) By weight, 11 parts of Mikania micrantha fermentation bacteria powder, 4 parts of bream larvae powder, 15 parts of calcium hydrogen phosphate, 5 parts of potassium aluminum sulfate dodecahydrate, 89 parts of 0.07 mol / L disodium hydrogen phosphate solution, and 73 parts of 0.83 mol / L sodium dihydrogen phosphate solution were mixed to obtain Mikania micrantha fermentation bacteria solution.

[0122] (9) By weight, 17 parts of ferrous chloride, 37.5 parts of magnesium sulfate, 14.7 parts of manganese chloride, 0.3 parts of γ-aminobutyric acid, 21 parts of sodium tetraborate decahydrate, 174 parts of lotus root starch, and 1000 parts of purified water were mixed to obtain a liquid fermentation medium for Mikania micrantha.

[0123] (10) Take 128 parts of Mikania micrantha to be fermented powder, 100 parts of Mikania micrantha fermentation liquid, and 818 parts of Mikania micrantha liquid fermentation medium by weight. Mix the Mikania micrantha to be fermented powder and Mikania micrantha fermentation liquid evenly and inoculate them on the Mikania micrantha liquid fermentation medium in the fermentation tank. Maintain the temperature in the tank at 14°C and exhaust once every 6 hours. Add 30 parts of Mikania micrantha fermentation liquid and 10 parts of corn flour by weight every day for 7 days. At 4:00, 10:00, 16:00, and 22:00 on the fourth day of fermentation, add 1 / 4 of the amount each time, for a total of 26 parts of alfalfa powder. At the same time, add 7 parts of kudzu powder every 2 days and 50 parts of fermented palm meal that has passed through an 80-mesh sieve 100% every 3 days. On the last day, put the tank aside and then add 16 parts of ammonium chloride and mix well. The mixture was centrifuged at 12000 r / min for 5 min at 14° C., the supernatant was discarded, and the precipitate was dried in an oven at 100° C. to obtain the dry powder of Mikania micrantha for aquatic feed of this embodiment.

[0124] Example 2

[0125] (1) By weight, 46 parts of fresh, unrotten Mikania micrantha stems and 14 parts of fresh, unrotten Mikania micrantha leaves were taken, dried in the dark at 22° C., and crushed until 100% passed through a 180-mesh sieve to obtain Mikania micrantha stem and leaf powder.

[0126] (2) By weight, 94 parts of Mikania micrantha stem and leaf powder, 4 parts of cellulase with an enzyme activity of 18650 U, and 631 parts of 0.05 mol / L sodium bicarbonate solution with a pH of 8.1 were mixed. The mixture was shaken at 34°C and 60 rpm for 60 h. The mixture was then centrifuged at 20°C and 5200 rpm for 8 min. The supernatant was collected to obtain Mikania micrantha enzymatic hydrolyzate.

[0127] (3) Mix 400 parts of Mikania micrantha enzymatic hydrolysate, 47 parts of 0.3 mol / L sodium hydroxide solution, and 2.8 parts of 0.8 mol / L sodium sulfite solution by weight. Treat at 120°C and 140 r / min for 2 h, then cool to room temperature. After adjusting the pH to 7.00, evaporate under reduced pressure at 70°C for 6 min, then cool to 7°C and remove surface crystals to obtain a Mikania micrantha slurry.

[0128] (4) Take 110 parts of Mikania micrantha thick paste, 24 parts of sodium alginate, and 1.35 parts of urea by weight, mix them evenly, freeze-dry at -90°C, and grind at low temperature until 100% passes through a 60-mesh sieve to obtain Mikania micrantha powder to be fermented.

[0129] (5) Place sterile, empty-bellied cotton aphids (Aphididae) on fresh, unrotten leaves of healthy Mikania plants at a density of 1 head:4 cm² and cover with perforated, colorless, transparent bags. The aphids are then sterilely cultured for 4 days at a temperature of 22°C, a light intensity of 15440 lux, and a photoperiod of L:D = 13 h:11 h. On the final day, the bags are removed, and all the insects are removed and crushed into a paste to obtain Mikania aphid paste.

[0130] (6) 198 parts by weight of Mikania micrantha aphid paste, 61 parts by weight of anhydrous ethanol, 3 parts by weight of n-hexane, and 20 parts by weight of gluten were placed in a fermentation tank. Fermentation was carried out at 25°C for 18 hours, followed by venting for 7 minutes every 3 hours. The fermentation tank was then opened after 55 hours. The fermented material was allowed to stand at room temperature for 7 hours, then dried in the dark at 13°C. The material was then ground into powder until 100% of the powder passed through an 80-mesh sieve to obtain Mikania micrantha fermentation powder.

[0131] (7) Take newly hatched bream fry, crush them into a paste, dry them at 103° C., grind them into powder at room temperature until 90% of the powder passes through an 80-mesh sieve, and obtain bream fry powder.

[0132] (8) By weight, 18 parts of Mikania micrantha fermentation bacteria powder, 6 parts of bream larvae powder, 17 parts of calcium hydrogen phosphate, 9 parts of potassium aluminum sulfate dodecahydrate, 96 parts of 0.07 mol / L disodium hydrogen phosphate solution, and 84 parts of 0.83 mol / L sodium dihydrogen phosphate solution were mixed to obtain Mikania micrantha fermentation bacteria liquid.

[0133] (9) By weight, 24 parts of ferrous chloride, 41.5 parts of magnesium sulfate, 19 parts of manganese chloride, 0.5 parts of γ-aminobutyric acid, 25.5 parts of sodium tetraborate decahydrate, 182 parts of lotus root starch, and 1100 parts of purified water were mixed to obtain a liquid fermentation medium for Mikania micrantha.

[0134] (10) Take 245 parts of Mikania micrantha to be fermented powder, 105 parts of Mikania micrantha fermentation liquid, and 1003 parts of Mikania micrantha liquid fermentation medium by weight. Mix the Mikania micrantha to be fermented powder and Mikania micrantha fermentation liquid evenly and inoculate them on the Mikania micrantha liquid fermentation medium in the fermentation tank. Maintain the temperature in the tank at 17°C and exhaust once every 7 hours. Add 39 parts of Mikania micrantha fermentation liquid and 13 parts of corn flour by weight every day for 12 days. At 4:00, 10:00, 16:00, and 22:00 on the fourth day of fermentation, add 1 / 4 of the amount each time, for a total of 28 parts of alfalfa powder. At the same time, add 8 parts of kudzu powder every 2 days and 53 parts of fermented palm meal that has passed through an 80-mesh sieve 100% every 3 days. On the last day, put the tank aside and then add 17 parts of ammonium chloride and mix well. The mixture was centrifuged at 14,000 r / min for 7 min at 18° C., the supernatant was discarded, and the precipitate was dried in an oven at 103° C. to obtain the dry powder of Mikania micrantha for aquatic feed of this embodiment.

[0135] Example 3

[0136] (1) By weight, 60 parts of fresh, unrotten Mikania micrantha stems and 16 parts of fresh, unrotten Mikania micrantha leaves were taken, dried in the dark at 27° C., and crushed until 100% passed through a 180-mesh sieve to obtain Mikania micrantha stem and leaf powder.

[0137] (2) By weight, 115 parts of Mikania micrantha stem and leaf powder, 5 parts of cellulase with an enzyme activity of 21,500 U, and 702 parts of 0.05 mol / L sodium bicarbonate solution with a pH of 8.2 were mixed. The mixture was shaken at 43°C and 60 rpm for 72 h. The mixture was then centrifuged at 25°C and 6000 rpm for 10 min. The supernatant was collected to obtain Mikania micrantha enzymatic hydrolyzate.

[0138] (3) Mix 502 parts of Mikania micrantha enzymatic hydrolysate, 55 parts of 0.4 mol / L sodium hydroxide solution, and 3.9 parts of 0.9 mol / L sodium sulfite solution by weight. Treat at 130°C and 150 rpm for 3 h, then cool to room temperature. Adjust the pH to 7.05, evaporate under reduced pressure at 80°C for 7 min, then cool to 10°C and remove surface crystals to obtain a Mikania micrantha slurry.

[0139] (4) Take 90-120 parts of Mikania micrantha thick paste, 21-27 parts of sodium alginate, and 0.14-2.22 parts of urea by weight, mix well, freeze-dry at -120 to -80°C, and grind at low temperature until 100% passes through a 60-mesh sieve to obtain Mikania micrantha powder to be fermented.

[0140] (5) Place sterile, empty-bellied cotton aphids (Aphididae) on fresh, unrotten leaves of healthy Mikania plants at a density of 2:3 cm² and place them in a perforated, colorless, transparent bag. The aphids are then sterilely cultured for 5 days at 24°C, 20,000 lux, and a photoperiod of L:D = 14 h:10 h. On the final day, remove the bag, remove all the insects, and crush them into a paste to obtain Mikania aphid paste.

[0141] (6) 228 parts by weight of Mikania micrantha aphid paste, 72 parts by weight of anhydrous ethanol, 6 parts by weight of n-hexane, and 28 parts by weight of gluten were placed in a fermentation tank. After fermentation at 30°C for 18 hours, the mixture was vented every 3 hours for 10 minutes. The fermentation tank was then opened after 60 hours. The fermented mixture was allowed to stand at room temperature for 8 hours, then dried in the dark at 15°C. The mixture was then ground into powder until 100% of the powder passed through an 80-mesh sieve to obtain Mikania micrantha fermentation powder.

[0142] (7) Take newly hatched bream fry, crush them into a paste, dry them at 105° C., grind them into powder at room temperature until 90% of the powder passes through an 80-mesh sieve, and obtain bream fry powder.

[0143] (8) By weight, 23 parts of Mikania micrantha fermentation bacteria powder, 8 parts of bream fry powder, 19 parts of calcium hydrogen phosphate, 12 parts of potassium aluminum sulfate dodecahydrate, 104 parts of 0.07 mol / L disodium hydrogen phosphate solution, and 95 parts of 0.83 mol / L sodium dihydrogen phosphate solution were mixed to obtain Mikania micrantha fermentation bacteria liquid.

[0144] (9) By weight, 33 parts of ferrous chloride, 45.5 parts of magnesium sulfate, 24.3 parts of manganese chloride, 0.7 parts of γ-aminobutyric acid, 30 parts of sodium tetraborate decahydrate, 191 parts of lotus root starch, and 1200 parts of purified water were mixed to obtain a liquid fermentation medium for Mikania micrantha.

[0145] (10) Take 360 ​​parts of Mikania micrantha to be fermented powder, 110 parts of Mikania micrantha fermentation liquid, and 1205 parts of Mikania micrantha liquid fermentation medium by weight. Mix the Mikania micrantha to be fermented powder and Mikania micrantha fermentation liquid evenly and inoculate them on the Mikania micrantha liquid fermentation medium in the fermentation tank. Maintain the temperature in the tank at 20°C and exhaust once every 8 hours. Add 50 parts of Mikania micrantha fermentation liquid and 16 parts of corn flour by weight every day for 16 days. At 4:00, 10:00, 16:00, and 22:00 on the fourth day of fermentation, add 1 / 4 of the amount each time, for a total of 32 parts of alfalfa powder. At the same time, add 9 parts of kudzu powder every 2 days and 55 parts of fermented palm meal that has passed through an 80-mesh sieve 100% every 3 days. On the last day, put the tank aside and then add 19 parts of ammonium chloride and mix well. The mixture was centrifuged at 15,000 r / min for 8 min at 20° C., the supernatant was discarded, and the precipitate was dried in an oven at 105° C. to obtain the dry powder of Mikania micrantha for aquatic feed of this embodiment.

[0146] Examples 4 to 6

[0147] The preparation is as follows: 28 parts of white fish meal, 41 parts of micrantha dry powder, 20 parts of α-starch, 0.1 part of choline chloride, 0.1 part of calcium hydrogen phosphate, 0.2 part of fish oil, 0.2 part of compound additives, 0.2 part of compound vitamins, 0.2 part of compound minerals, and 0.1 part of adhesive. The compound additives are prepared by mixing the following raw materials in parts by weight: 12 parts of β-glucanase with an enzyme activity of 78,000 U / g, 9 parts of β-xylosidase with an enzyme activity of 228,000 U / g, 35 parts of β-D-glucose oxidoreductase with an enzyme activity of 6,730 U / g, 5 parts of α-galactosidase with an enzyme activity of 7,500 U / g, 6 parts of bream fry meal, and 14 parts of micrantha fermented bacteria powder. The multivitamin is prepared by mixing the following raw materials in parts by weight: 22 parts vitamin A, 40 parts vitamin C, 0.2 parts riboflavin, 0.07 parts folic acid, 0.03 parts vitamin B12, 0.3 parts biotin, 125 parts vitamin D3, 12 parts thiamine, 32 parts vitamin K, 7 parts vitamin E, 22 parts vitamin B6, 80 parts inositol, 15 parts pantothenic acid, and 63 parts niacinamide. The multimineral is prepared by mixing the following raw materials in parts by weight: 15 parts zinc chloride, 4 parts calcium iodate, 0.2 parts potassium fluoride, 12 parts copper sulfate, 0.14 parts sodium molybdate, 137 parts manganese chloride, 172 parts magnesium sulfate, 1 part sodium selenite, 11.3 parts cobalt sulfate, and 330 parts ferrous sulfate.

[0148] Among them, the dry powder of Mikania micrantha in Examples 1 to 3 is respectively prepared to obtain Examples 4 to 6. The preparation method is as follows: all solid feed raw materials are crushed with a grinder and completely passed through a 120-mesh standard sieve. Take choline chloride, add calcium hydrogen phosphate, adhesive, compound additives, compound vitamins, α-starch, white fish meal, Mikania micrantha dry powder, fish oil, and water in sequence, and mix thoroughly after each raw material is added. Finally, mix thoroughly and evenly. Pour all the above feed raw materials into a pelletizer according to proportion and process for 10 minutes. The moisture content of the feed is 20% based on wet weight. The feed is finally made into columnar pellets with a diameter: height ratio of 1:9 and a diameter of 1 / 10 of the eel's mouth crack through the discharge port.

[0149] Examples 7 to 9

[0150] The preparation is as follows: 32 parts white fish meal, 44 parts micrantha dry powder, 23 parts α-starch, 0.2 parts choline chloride, 0.2 parts calcium hydrogen phosphate, 0.4 parts fish oil, 0.3 parts composite additives, 0.3 parts composite vitamins, 0.3 parts composite minerals, and 0.2 parts adhesive. The composite additives are prepared by mixing the following raw materials in parts by weight: 14 parts β-glucanase with an enzyme activity of 96,000 U / g, 13 parts β-xylosidase with an enzyme activity of 290,000 U / g, 44 parts β-D-glucose oxidoreductase with an enzyme activity of 8,260 U / g, 16 parts α-galactosidase with an enzyme activity of 8,550 U / g, 11 parts bream fry meal, and 22 parts micrantha fermented bacteria powder. The multivitamin is prepared by mixing the following raw materials in parts by weight: 26 parts vitamin A, 52 parts vitamin C, 0.31 parts riboflavin, 0.1 parts folic acid, 0.05 parts vitamin B12, 0.6 parts biotin, 180 parts vitamin D3, 15 parts thiamine, 33 parts vitamin K, 9 parts vitamin E, 25 parts vitamin B6, 90 parts inositol, 22 parts pantothenic acid, and 70 parts niacinamide. The multimineral is prepared by mixing the following raw materials in parts by weight: 17 parts zinc chloride, 11 parts calcium iodate, 0.3 parts potassium fluoride, 13 parts copper sulfate, 0.16 parts sodium molybdate, 140 parts manganese chloride, 187 parts magnesium sulfate, 1.1 parts sodium selenite, 11.7 parts cobalt sulfate, and 347 parts ferrous sulfate.

[0151] Among them, the dry powder of Mikania micrantha in Examples 1 to 3 is respectively prepared to obtain Examples 7 to 9. The preparation method is as follows: all solid feed raw materials are crushed with a grinder and completely passed through a 120-mesh standard sieve. Take choline chloride, add calcium hydrogen phosphate, adhesive, compound additives, compound vitamins, α-starch, white fish meal, Mikania micrantha dry powder, fish oil, and water in sequence, and mix thoroughly after each raw material is added. Finally, mix thoroughly and evenly. Pour all the above feed raw materials into a pelletizer according to proportion and process for 15 minutes. The moisture content of the feed is 25.5% based on wet weight. The feed is finally made into columnar pellets with a diameter: height ratio of 2:7 and a diameter of 1 / 9 of the eel's mouth crack through the discharge port.

[0152] Examples 10 to 12

[0153] The preparation is as follows: 34 parts of white fish meal, 47 parts of micrantha dry powder, 25 parts of α-starch, 0.3 parts of choline chloride, 0.3 parts of calcium hydrogen phosphate, 0.5 parts of fish oil, 0.4 parts of compound additives, 0.4 parts of compound vitamins, 0.4 parts of compound minerals, and 0.3 parts of adhesive. The compound additives are prepared by mixing the following raw materials in parts by weight: 15 parts of β-glucanase with an enzyme activity of 130,000 U / g, 17 parts of β-xylosidase with an enzyme activity of 360,000 U / g, 56 parts of β-D-glucose oxidoreductase with an enzyme activity of 10,800 U / g, 26 parts of α-galactosidase with an enzyme activity of 9,600 U / g, 16 parts of bream fry meal, and 28 parts of micrantha fermented bacteria powder. The multivitamin is prepared by mixing the following raw materials in parts by weight: 30 parts vitamin A, 65 parts vitamin C, 0.45 parts riboflavin, 0.18 parts folic acid, 0.06 parts vitamin B12, 0.9 parts biotin, 238 parts vitamin D3, 17 parts thiamine, 34 parts vitamin K, 11 parts vitamin E, 28 parts vitamin B6, 100 parts inositol, 29 parts pantothenic acid, and 74 parts niacinamide. The multimineral is prepared by mixing the following raw materials in parts by weight: 19 parts zinc chloride, 15 parts calcium iodate, 0.5 parts potassium fluoride, 14 parts copper sulfate, 0.18 parts sodium molybdate, 142 parts manganese chloride, 190 parts magnesium sulfate, 1.2 parts sodium selenite, 12 parts cobalt sulfate, and 365 parts ferrous sulfate.

[0154] Among them, the dry powder of Mikania micrantha of Examples 1 to 3 is respectively prepared to obtain Examples 10 to 12. The preparation method is as follows: all solid feed raw materials are crushed with a grinder and completely passed through a 120-mesh standard sieve. Take choline chloride, add calcium hydrogen phosphate, adhesive, compound additives, compound vitamins, α-starch, white fish meal, Mikania micrantha dry powder, fish oil, and water in sequence, and mix thoroughly after each raw material is added. Finally, mix thoroughly and evenly. Pour all the above feed raw materials into a pelletizer according to proportion and process for 20 minutes. The moisture content of the feed is 35% based on wet weight. The feed is finally made into columnar pellets with a diameter: height = 1:2 and a diameter of 1 / 7 of the eel's mouth through the discharge port.

[0155] Test example

[0156] (1) Test method

[0157] (1.1) Test conditions

[0158] Before the experiment began, the aquaculture tanks were cleaned and filled with water. Formaldehyde was evenly sprinkled into the tanks and allowed to soak for 24 hours to disinfect them. The aquaculture water was tap water that had been stagnant for 24 hours and had been aerated with an air pump to maintain a dissolved oxygen level of 10 mg / L. The water temperature was naturally maintained between 24 and 29°C, and the pH was between 7.4 and 9.2.

[0159] (1.2) Test preparation

[0160] Several adult eels weighing 100 to 150 g from the same batch that have been acclimated to eating artificial feed and several broodstock eels weighing 600 to 800 g from the same batch that have been acclimated to eating artificial feed were selected and temporarily cultured in an indoor recirculating aquaculture system for 2 weeks. The aquatic feed provided in Example 8 was replaced with an equal amount of white fish meal using micrantha dry powder, and the composite additive was replaced with an equal amount of adhesive. The eels were fed according to a satiation amount to allow them to gradually adapt to the feed and the culture environment.

[0161] Adult eel group: Healthy and uniform-sized eels (138.76±8.11 g) were randomly divided into nine experimental groups (experimental groups 1 to 9, 6 replicates per group) and three control groups (control groups 1, 2, and 3, 6 replicates per group). The eels were placed in cylindrical plastic aquariums (volume: 2 m3) in an indoor recirculating aquaculture system. 3 ), the stocking density is 30 tails / tank (about 2kg / m 3 ).

[0162] Broodstock group: Healthy and uniform-sized eels (727.21±10.16 g) were randomly divided into two experimental groups (experimental groups 10 and 11, 6 replicates in each group) and two control groups (control groups 4 and 5, 6 replicates in each group). They were placed in cylindrical cement culture ponds (volume: 5 m3) in an indoor recirculating aquaculture system. 3 ), the stocking density is tail / pond (about 1.2kg / m 3 , half male and half female).

[0163] (1.3) Feeding mode

[0164] Experimental groups 1 to 9 were fed with the eel feed provided in Examples 4 to 12 respectively;

[0165] Experimental group 10: fed with the eel feed provided in Example 4;

[0166] Experimental group 11: fed with the eel feed provided in Example 12;

[0167] Control group 1: fed with the feed of Example 8, except that the dry powder of Mikania micrantha in the aquatic feed was replaced with white fish meal of equal protein content;

[0168] Control group 2: fed with the feed of Example 8, except that the dry powder of Mikania micrantha in aquatic feed was replaced with an equal amount of Mikania micrantha fermented powder;

[0169] Control group 3: fed with the feed of Example 8, except that the composite additive was replaced with an equal amount of bentonite;

[0170] Control group 4: fed with the feed of Example 8, except that the dry powder of Mikania micrantha in the aquatic feed was replaced with white fish meal of the same protein content;

[0171] Control group 5: fed with the feed of Example 8, except that the dry powder of Mikania micrantha in the aquatic feed was replaced with corn flour with the same glucose content;

[0172] During the formal experimental phase, the average daily protein intake per tank during the experimental preparation period was used to ensure that the protein intake of each group of eels and broodstock was equal (but not equal for the adult and broodstock groups). The fish were fed three times daily at designated locations and times, with feeding times controlled between 9:00 AM and 10:00 AM, 3:00 PM and 4:00 PM, and 8:00 PM and 9:00 PM. After feeding, feces, leftover bait, and other debris were removed from the aquaculture environment using a suction device. The water temperature and pH of the aquaculture environment were monitored. The food intake of each group of fish was observed and recorded daily during the feeding period.

[0173] (1.4) Daily management

[0174] (1.4.1) Daily management of adult eels

[0175] Before and after the experiment, the weight of all eels in each experimental group was counted to calculate body weight gain. This experiment used an intelligent, fully automatic circulating water system to control the aquaculture water environment. Every afternoon at 4:00 PM, the activity of the experimental fish was observed and water quality conditions, such as dissolved oxygen, temperature, and pH, were recorded to ensure the experimental fish lived in a good water quality environment. Daily aquaculture records were kept, and any dead fish were promptly disposed of and counted. The experimental period lasted eight weeks. At the end of the experiment, six eels whose weight was within the final average weight within the tank ±10g were randomly selected from each tank to test the content of relevant substances and growth indicators.

[0176] (1.4.2) Daily management of broodstock group

[0177] This experiment uses an intelligent fully automatic circulating water system to control the aquaculture water environment. Every afternoon at 4:00 PM, the activity of the experimental fish was observed and the water quality conditions such as dissolved oxygen, water temperature, and pH of the aquaculture water were recorded to ensure that the experimental fish lived in a good water quality environment. The experimental period was 25 weeks. After completing the aquaculture experiment from Weeks 1 to 8, the total weight of all groups was calculated; then, 4 eels (half male and half female) with a final average weight in the tank of ±30g were randomly selected from each pond and their gonads were dissected and used to calculate the gonad index (the last day of Week 8). In Week 9, all groups were treated identically to complete the induced spawning experiment, and the survival rate of the broodstock after induced spawning was calculated (conducted and completed on Day 3 of Week 9). The hatching operation of fertilized eggs in each group was completed at the same time in the 9th week. Then 200 willow leaf eels were selected from each group (day 1 of the 10th week). After 16 weeks of culture under the same conditions (DO = 12 mg / L, T = 23℃, pH = 8), the deformity rate of the newly hatched fry in the willow leaf eel stage was calculated in groups on the last day of the 25th week.

[0178] (2) Test results

[0179] (2.1) Comparison of toxic substance content in whole eel of adult eel group.

[0180] The toxic substance contents of whole eels in each adult eel group were detected, and the results are shown in Table 1. The contents in Table 1 are dry weight, unit: %. The data in Table 1 are shown as mean ± standard error; the differences between different shoulder marks are significant (P<0.05).

[0181] Table 1

[0182]

[0183] As shown in Table 1, eels fed a feed containing the Mikania micrantha dry powder for aquatic feed according to the present invention had significantly lower levels of toxic substances such as sesquiterpene lactones, volatile oils, and ketones than eels fed a feed containing untreated Mikania micrantha fermented powder. This indicates that the Mikania micrantha dry powder for aquatic feed according to the present invention has extremely low toxicity and that the preparation process is highly effective in detoxifying Mikania micrantha.

[0184] (2.2) Comparison of some nutritional components and growth indicators of adult eels.

[0185] The total amino acid content, reducing sugar content, feed efficiency, feeding rate, weight gain rate, protein efficiency ratio, protein retention rate, and specific growth rate of adult eels were analyzed and statistically analyzed, all calculated on a dry weight basis. The results are shown in Table 2. The data in Table 2 are presented as mean ± standard error; significant differences were observed between groups with different shoulder marks (P < 0.05).

[0186] Table 2

[0187]

[0188] As shown in Table 2, in terms of nutrient-related indicators: the total amino acid content and reducing sugar content of eels fed with feed containing dry powder of Mikania micrantha for aquatic feed were higher than those of eels fed with unreplaced white fish meal, eels fed with fish meal replaced by fermented powder of Mikania micrantha for aquatic feed, and eels fed with fish meal replaced by corn meal.

[0189] In terms of growth indicators: the dry powder of Mikania micrantha for aquatic feed of the present invention has the advantage of replacing white fish meal in eel feed in terms of improving feed efficiency, feeding rate, weight gain rate, protein efficiency ratio, protein retention rate and specific growth rate, and also has advantages over unfermented Mikania micrantha to be fermented powder.

[0190] It can be seen that the composite additive in the eel feed of the present invention also has the effect of improving feed efficiency, feeding rate, weight gain rate, protein efficiency ratio, protein retention rate and specific growth rate.

[0191] (2.3) Comparison of some artificial propagation indicators of eels and their larvae and juveniles in the broodstock group.

[0192] The gonadal index, post-induced broodstock survival rate, and deformity rate of newly hatched fry in the eel stage of each broodstock group were statistically analyzed, and the results are shown in Table 3. The data in Table 3 are presented as mean ± standard error; the differences between the different shoulder marks were significant (P < 0.05).

[0193] Table 3

[0194] Experimental group 10 Experimental group 11 Control group 4 Control group 5 Gonadal index, % 8.21±0.00b 9.23±1.63c 8.27±0.04b 6.40±0.58a Broodstock survival rate, % 76.43±4.22b 82.65±2.22b 75.16±3.65b 61.00±4.30a Deformity rate of eels, % 23.47±1.20a 22.03±0.32a 30.91±0.04b 32.74±0.59b

[0195] As shown in Table 3, the dry powder of Mikania micrantha for aquatic feed of the present invention, as the proportion of fish meal in the eel feed replaced by the dry powder is different, the eels in the test group have great numerical advantages in terms of gonad index, survival rate of broodstock after induced spawning, and deformity rate of newly hatched fry in the eel stage, compared with the control group 5 which fed the same energy feed with corn meal instead of fish meal.

[0196] Comparing the control group 4 that fed on feed without fish meal replacement with the test groups 4 and 5, the dry powder of Mikania micrantha for aquatic feed of the present invention had a numerical advantage close to that of the eels in the control group 4 that fed on feed with corn meal instead of fish meal, which is also an energy feed, and was even slightly better than that of the eels in the control group 4.

[0197] In summary, combined with the performance in Tables 1 to 3, it can be seen that: 1) the toxicity of the dry powder of Mikania micrantha for aquatic feed of the present invention is extremely low, and the preparation process has an excellent detoxification effect on Mikania micrantha; 2) the dry powder of Mikania micrantha for aquatic feed of the present invention is a high-quality choice for replacing white fish meal in eel feed; 3) in terms of replacing white fish meal, the dry powder of Mikania micrantha for aquatic feed of the present invention is superior to corn flour, which is also an energy feed; 4) the dry powder of Mikania micrantha for aquatic feed of the present invention and the eel feed containing the dry powder can improve the gonad index of eels during artificial reproduction and the survival rate of broodstock after induced spawning, reduce the deformity rate of newly hatched fry in the willow leaf eel stage, and at the same time increase the total amino acid content and reducing sugar content of the whole eel during artificial breeding and rearing, and improve the feed efficiency, feeding rate, weight gain rate, protein efficiency ratio, protein retention rate and specific growth rate of eels.

[0198] Although the present invention has been illustrated and described using specific embodiments, it should be appreciated that the above embodiments are merely intended to illustrate the technical solutions of the present invention rather than to limit them. Those skilled in the art should understand that the technical solutions described in the above embodiments may be modified, or some or all of the technical features thereof may be replaced by equivalents, without departing from the spirit and scope of the present invention. However, these modifications or replacements do not deviate from the essence of the corresponding technical solutions within the scope of the technical solutions of the embodiments of the present invention. Therefore, this means that all such replacements and modifications within the scope of the present invention are included in the appended claims.

Claims

1. A method for preparing micrantha dry powder, characterized in that: The steps include: (1) preparing fresh Mikania micrantha stems and fresh Mikania micrantha leaves, drying, crushing, and sieving to obtain Mikania micrantha stem and leaf powder; then fully mixing the Mikania micrantha stem and leaf powder, cellulase, and sodium bicarbonate solution, and performing a first reaction to separate the solid and liquid to obtain Mikania micrantha hydrolyzate; fully mixing the Mikania micrantha hydrolyzate, sodium hydroxide solution, and sodium sulfite solution, and performing a second reaction to adjust the pH to 6.9-7.1, and then performing reduced pressure evaporation, cooling, and removing surface crystals to obtain Mikania micrantha thick paste; fully mixing the Mikania micrantha thick paste, sodium alginate, and urea, and performing freeze-drying, crushing, and sieving to obtain Mikania micrantha fermentation powder; (2) placing adult cotton aphids on fresh Mikania micrantha leaves and aseptically cultivating them for 2 to 5 days, then crushing the insect bodies to obtain Mikania micrantha aphid paste; mixing the Mikania micrantha aphid paste, anhydrous ethanol, n-hexane, and gluten for fermentation, and after the fermentation is completed, standing, drying in the dark, crushing, and sieving to obtain Mikania micrantha fermentation powder; (3) preparing the micrantha fermentation bacteria powder, bream larvae powder, calcium hydrogen phosphate, potassium aluminum sulfate dodecahydrate, disodium hydrogen phosphate solution and sodium dihydrogen phosphate solution, and fully mixing to obtain the micrantha fermentation bacteria liquid; (4) preparing an aqueous solution of ferrous chloride, magnesium sulfate, manganese chloride, γ-aminobutyric acid, sodium tetraborate decahydrate, and lotus root starch as a liquid fermentation medium for Mikania micrantha; (5) The micrantha to-be-fermented powder and the micrantha fermentation broth are uniformly mixed, inoculated into the micrantha liquid fermentation medium and fermented, and the micrantha fermentation broth, corn flour, alfalfa powder, kudzu powder, and fermented palm meal are regularly added; after the fermentation is completed, the tank is placed, ammonium chloride is added to the fermentation product and the mixture is fully mixed, and then a solid phase is obtained by solid-liquid separation, and the moisture of the solid phase is removed to obtain the micrantha dry powder.

2. The method for preparing micrantha dry powder according to claim 1, wherein Step (1) includes at least one of the following features (a) to (f): (a) the mass ratio of the fresh Mikania micrantha stems to the fresh Mikania micrantha leaves is (30-60):(10-16); (b) the mass ratio of the Mikania micrantha stem and leaf powder, the cellulase and the sodium bicarbonate solution is (84-115): (3-5): (525-702); (c) the mass ratio of the Mikania micrantha enzymatic hydrolysate, the sodium hydroxide solution and the sodium sulfite solution is (286-502): (33-55): (1.7-3.9); (d) the mass ratio of the micrantha micrantha slurry, the sodium alginate and the urea is (90-120):(21-27):(0.14-2.22); (e) The conditions of the first reaction include: a temperature of 28° C. to 43° C. and a duration of 48 h to 72 h; (f) The conditions of the second reaction include: temperature of 110° C. to 130° C., and duration of 1.8 h to 3 h.

3. The method for preparing micrantha dry powder according to claim 1, wherein: Step (2) includes at least one of the following features (a) to (c): (a) The sterile culture conditions include: a temperature of 18°C ​​to 24°C, a light intensity of 10,000 lux to 20,000 lux, and a light duration L:D of (10 to 14) h:(10 to 14) h; (b) the mass ratio of the Mikania micrantha aphid paste, the anhydrous ethanol, the n-hexane and the gluten is (160-228): (34-72): (1-6): (13-28); (c) The fermentation conditions include: a temperature of 22° C. to 30° C., a fermentation time of 48 to 60 hours, and exhaust gas for 5 to 10 minutes every 3 hours after the 18th hour of the fermentation.

4. The method for preparing micrantha dry powder according to claim 1, wherein In step (3), the fermented bacteria liquid of Mikania micrantha comprises the following components in parts by weight: 11-23 parts of the fermented bacteria powder of Mikania micrantha, 4-8 parts of the bream fry powder, 15-19 parts of the calcium hydrogen phosphate, 5-12 parts of the potassium aluminum sulfate dodecahydrate, 89-104 parts of the disodium hydrogen phosphate solution, and 73-95 parts of the sodium dihydrogen phosphate solution; And / or, in step (4), the aqueous solution of the liquid fermentation medium of Mikania micrantha includes the following components in parts by weight: 17 to 33 parts of ferrous chloride, 37.5 to 45.5 parts of magnesium sulfate, 14.7 to 24.3 parts of manganese chloride, 0.3 to 0.7 parts of γ-aminobutyric acid, 21 to 30 parts of sodium tetraborate decahydrate, 174 to 191 parts of lotus root starch, and 1000 to 1200 parts of water.

5. The method for preparing micrantha dry powder according to claim 1, characterized in that: In step (5), 128 to 360 parts of the Mikania micrantha to be fermented powder and 100 to 110 parts of the Mikania micrantha fermentation liquid are mixed evenly by mass, and the mixture is inoculated into 818 to 1205 parts of the Mikania micrantha liquid fermentation medium; after the fermentation is completed, the tank is placed, and 16 to 19 parts of ammonium chloride are added to the fermentation product and mixed thoroughly; And / or, the fermentation culture conditions include: a temperature of 14° C. to 20° C., a duration of 7 to 16 days; and exhaust every 6 to 8 hours.

6. The Mikania micrantha dry powder obtained by the method for preparing Mikania micrantha dry powder according to any one of claims 1 to 5.

7. An eel feed, characterized in that: The composition comprises the following components in parts by weight: 41-47 parts of the dry powder of Mikania micrantha according to claim 6; and 28-34 parts of white fish meal, 20-25 parts of alpha-starch, 0.1-0.3 parts of choline chloride, 0.1-0.3 parts of calcium hydrogen phosphate, 0.2-0.5 parts of fish oil, 0.2-0.4 parts of compound additives, 0.2-0.4 parts of compound vitamins, 0.2-0.4 parts of compound minerals, and 0.1-0.3 parts of adhesive.

8. The eel feed according to claim 7, characterized in that Includes at least one of the following features (A) to (C): (A) The composite additive comprises the following components by mass: 12 to 15 parts of β-glucanase, 9 to 17 parts of β-xylosidase, 35 to 56 parts of β-D-glucose oxidoreductase, 5 to 26 parts of α-galactosidase, 6 to 16 parts of bream fry meal, and 14 to 28 parts of the fermented bacteria powder of Mikania micrantha; (B) the vitamin complex comprises the following ingredients, calculated by mass: 22-30 parts of vitamin A, 40-65 parts of vitamin C, 0.2-0.45 parts of riboflavin, 0.07-0.18 parts of folic acid, 0.03-0.06 parts of vitamin B1, 0.3-0.9 parts of biotin, 125-238 parts of vitamin D3, 12-17 parts of thiamine, 32-4 parts of vitamin K, 7-11 parts of vitamin E, 22-28 parts of vitamin B6, 80-100 parts of inositol, 15-29 parts of pantothenic acid, and 63-74 parts of niacinamide; (C) The composite mineral comprises the following components in parts by mass: 15-19 parts of zinc chloride, 4-15 parts of calcium iodate, 0.2-0.5 parts of potassium fluoride, 12-14 parts of copper sulfate, 0.14-0.18 parts of sodium molybdate, 137-142 parts of manganese chloride, 172-190 parts of magnesium sulfate, 1-1.2 parts of sodium selenite, 11.3-12 parts of cobalt sulfate and 330-365 parts of ferrous sulfate.

9. A method for preparing eel feed according to claim 7 or 8, characterized in that: The steps include: The solid phase components of the eel feed are crushed and sieved respectively, and then the components of the eel feed are fully mixed and granulated to obtain the eel feed.

10. Use of the dry powder of Mikania micrantha according to claim 6 and the eel feed according to claim 7 or 8 in aquaculture.

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