Environment-friendly compound feed for parent shrimps of litopenaeus vannamei and preparation method of environment-friendly compound feed

Abstract

The invention discloses an environment-friendly compound feed for parent shrimps of litopenaeus vannamei and a preparation method of the environment-friendly compound feed for the parent shrimps of the litopenaeus vannamei. Comprising the following raw materials in percentage by weight: 40% of fish meal, 14% of mussel powder, 13.75% of high gluten flour, 9.48% of krill powder, 7% of fish oil, 6% of casein, 2% of phospholipid, 1.52% of probiotics, 1.5% of monocalcium phosphate, 1% of choline, 0.85% of an additive for promoting gonad development, 0.75% of vitamin C, 0.5% of cholesterol, 0.5% of astaxanthin, 0.5% of composite vitamins, 0.5% of composite minerals and 0.15% of traditional Chinese medicinal materials. The feed raw materials are crushed and uniformly mixed by adopting a step-by-step amplification method, then water is added for stirring, the mixture is pressed into the parent shrimp feed in a granulator, and the parent shrimp feed is stored in a shady and cool place for later use after being dried. The method can ensure that the immunity and disease resistance of the parent shrimps are improved by feeding the parent shrimps with the compound feed, and meanwhile, the breeding water body of the parent shrimps is stable and lasting.

Description

Technical Field

[0001] The invention belongs to the field of aquatic animal nutrition and feed, and particularly relates to an environmentally friendly compound feed for broodstock of Penaeus vannamei and a preparation method thereof. Background Art

[0002] With the continuous development of my country's shrimp farming industry, artificially formulated broodstock feed has been developed, mainly through the following technologies: (i) by increasing the protein and energy levels in the feed formula, through high protein and high energy to meet the high energy consumption of broodstock during the breeding period. (ii) Optimizing the nutritional ratio: through scientific proportions, adding essential amino acids, fatty acids, vitamins and minerals, promoting the development of broodstock gonads, and improving the egg production and larval quality.

[0003] For example, the prior art: a feed for broodstock of Penaeus vannamei (CN201610677562.9) that can replace fresh bait is added with high-protein, high-energy raw materials - fish meal, krill meal, mussel meal and casein are added to the feed as protein sources and their proportions are adjusted, so that the feed has a reasonable nutritional component and can meet the nutritional needs of Penaeus vannamei broodstock for protein and amino acids.

[0004] Adding a certain proportion of krill meal, a certain amount of fish oil, cholesterol and astaxanthin to the feed of Penaeus vannamei broodstock can meet the needs of Penaeus vannamei broodstock for essential fatty acids and cholesterol, thereby increasing their egg production, hatching rate and metamorphosis rate.

[0005] Broodstock feed is mainly used to meet the special nutritional needs of white shrimp during the breeding period and to improve the breeding performance and larvae quality of broodstock.

[0006] Although the prior art has achieved certain results in the field of broodstock feed, there are still the following problems and shortcomings:

[0007] 1. Unbalanced gonadal development: Although the feed in the prior art can promote gonadal development, in actual application, the gonadal development speed of some broodstock is too fast, resulting in inconsistent ovarian maturity and affecting the breeding effect.

[0008] 2. Limited improvement in immunity and disease resistance: Although immune enhancers are added to the feed in the prior art, the effect of improving the immunity and disease resistance of broodstock is limited in the actual breeding process.

[0009] 3. Insufficient stability of breeding water: While promoting the reproduction of broodstock, the existing feed technology fails to fully consider the stability of the breeding water, which may lead to excessive accumulation of nutrients in the water and affect the ecological balance of the water.

[0010] 4. The quality of juveniles needs to be improved: Although the feeds in the existing technology can improve the quality of juveniles, in practical applications, the quality of juveniles still needs to be further improved.

[0011] The existence of these problems and shortcomings is mainly due to the fact that the feed formulas in the prior art are not scientific and reasonable enough in the screening and proportioning of nutrients, and the production process has limitations. Summary of the invention

[0012] The purpose of the present invention is to provide an environmentally friendly compound feed for broodstock of Penaeus vannamei and a preparation method thereof, by fermenting Chinese medicine with probiotics, combining additives that promote gonad development, screening out protein sources, amino acids, lipids, vitamins and mineral elements, and making a feed that can effectively solve the problems of the prior art. It is ensured that the immunity and disease resistance of broodstock are improved by feeding compound feed, and at the same time, the breeding water of broodstock is stable and durable; the ovaries of broodstock develop and mature quickly, the gonad index is improved, the egg carrying capacity is large, and the number and quality of nauplii are good, so as to solve the problems raised in the above background technology.

[0013] To achieve the above-mentioned purpose, the present invention provides the following technical scheme: an environmentally friendly compound feed for Penaeus vannamei broodstock, comprising the following raw materials in weight percentage ratio: 40% fish meal, 14% mussel powder, 13.75% high-gluten flour, 9.48% krill meal, 7% fish oil, 6% casein, 2% phospholipids, 1.52% probiotics, 1.5% monocalcium phosphate, 1% choline, 0.85% additives for promoting gonadal development, 0.75% vitamin C, 0.5% cholesterol, 0.5% astaxanthin, 0.5% complex vitamins, 0.5% complex minerals, and 0.15% Chinese medicinal materials.

[0014] Furthermore, the additive for promoting gonadal development is formulated in mg / kg feed, which includes the following raw materials in proportion by weight: 500 mg folic acid, 2000 mg soy isoflavones, 5000 mg krill meal, 800 mg lecithin, 200 mg progesterone, etc.

[0015] Furthermore, the compound vitamin is formulated in mg / kg feed, and includes the following raw materials in weight proportions: 25 mg thiamine; 45 mg riboflavin; 20 mg pyridoxine hydrochloride; 0.1 mg vitamin B12; 10 mg vitamin K3; 800 mg inositol; 60 mg pantothenic acid; 200 mg niacin; 120 mg biotin; 32 mg vitamin A; 5 mg vitamin D; 120 mg vitamin E; 3660 mg sub-powder.

[0016] Furthermore, the composite mineral is formulated in mg / kg feed, and includes the following raw materials in the following weight proportions: 2 mg sodium fluoride; 0.8 mg potassium iodide; 50 mg cobalt chloride; 10 mg copper sulfate; 80 mg iron sulfate; 50 mg zinc sulfate; 1200 g magnesium sulfate; 3000 mg calcium dihydrogen phosphate; and 1551 mg zeolite powder.

[0017] Furthermore, the probiotics are formulated in mg / kg feed ratio, which includes the following raw materials in the following weight proportions: 10 mg of Clostridium butyricum, 5 mg of yeast, and 10 mg of Bacillus subtilis.

[0018] Furthermore, the Chinese medicine is formulated in mg / kg feed, and includes the following raw materials in the following weight proportions: 5 mg of Astragalus, 6 mg of Atractylodes, 3.5 mg of Clove, 2.5 mg of Saposhnikovia and 12 mg of ginsenoside.

[0019] The present invention provides another technical solution: a method for preparing an environmentally friendly compound feed for Penaeus vannamei broodstock, and the specific preparation steps are as follows:

[0020] S1. Probiotics and Chinese medicine fermentation. The selected probiotics are: Clostridium butyricum, yeast, Bacillus subtilis. The selected Chinese medicines are: Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata and ginsenosides. The solid fermentation method is used to ferment Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata and ginsenosides, including raw material processing, strain preparation, fermentation material preparation, inoculation and fermentation, post-fermentation processing, and fermentation effect detection.

[0021] Furthermore, the specific method of raw material processing is as follows: (1) Crushing particle size: crush the Chinese medicine material to 60 mesh to ensure that the microorganisms can fully contact the Chinese medicine ingredients during the fermentation process and improve the fermentation efficiency. (2) Addition of ginsenoside: ginsenoside can be dissolved and then evenly sprayed into the wet Chinese medicine material to ensure its uniform distribution.

[0022] Furthermore, the specific method for preparing the bacterial strains is as follows: (1) Yeast: activated in a medium containing glucose at 30°C for 12 hours. (2) Clostridium butyricum: activated at 37°C for 24 hours under anaerobic conditions. (3) Bacillus subtilis: activated in a medium containing peptone and glucose at 30°C for 12 hours. (4) Bacterial strain ratio: the ratio of Clostridium butyricum, yeast, and Bacillus subtilis is 1:1:1 (calculated according to the bacterial solution concentration), which can be adjusted according to experimental results.

[0023] Furthermore, the specific method of preparing the fermentation material is as follows: (1) Water content control: ensure that the water content of the wet material is about 45%. Too high or too low a water content will affect the fermentation effect. (2) Nutrient addition: add 2-3 grams of glucose, 1-2 grams of peptone, and 0.1 grams of potassium dihydrogen phosphate to every 100 grams of Chinese medicine wet material to meet the growth requirements of the strain.

[0024] Furthermore, the specific method of inoculation and fermentation is as follows: (1) The inoculation amount is 10% (volume ratio) of the total amount of bacterial liquid in every 100 grams of wet material to ensure that the bacteria can reproduce rapidly. (2) Fermentation conditions are set: Early aerobic fermentation: after inoculation of yeast, aerobic fermentation is carried out at 30°C for 12 hours. Late anaerobic fermentation: after inoculation of Clostridium butyricum and Bacillus subtilis, anaerobic fermentation is carried out at 35°C for 48 hours.

[0025] Furthermore, the specific method of post-fermentation treatment is as follows: (1) Drying temperature: After fermentation is completed, the material is dried at 50°C to a moisture content of less than 10% to avoid high temperature damage to the active ingredients. (2) Crushing and sieving: The dried material is crushed to 100 mesh and sieved for later use.

[0026] Furthermore, the specific method of the fermentation effect detection is as follows: (1) Active ingredient detection: High performance liquid chromatography (HPLC) is used to detect the content of active ingredients such as astragalus polysaccharides and ginsenosides. (2) Pharmacological activity evaluation: The pharmacological activity of the fermentation product is evaluated through in vitro experiments (such as antioxidant experiments, immune cell activity experiments) and animal experiments.

[0027] S2. Prepare the raw materials according to the above weight percentages, and then mix and stir them evenly; crush the mixed raw materials through an ultrafine grinder so that all ingredients can pass through a 200-320um sieve;

[0028] S3, then move the material into the modem, pass 90-95°C steam into it for modulation for 5-10 minutes, and then extrude and granulate it at 85-90°C to form broodstock feed with a specification of 1.5×3.0mm;

[0029] S4. Dry the feed pellets at 80-85°C.

[0030] Compared with the prior art, the present invention has the following beneficial effects:

[0031] 1. The environmentally friendly compound feed for Penaeus vannamei broodstock and its preparation method add fish meal, krill meal, squid meal, fermented soybean meal and unicyclidine as protein sources to the feed and adjust their proportions, so that the feed has a reasonable nutritional component and can meet the nutritional needs of Penaeus vannamei broodstock for protein and amino acids. In addition, the addition of krill meal, unicyclidine and sea flavor can significantly improve the feeding appeal of shrimp feed.

[0032] 2. The environmentally friendly formula feed for Penaeus vannamei broodstock and the preparation method thereof, by adding a certain proportion of krill powder, a certain amount of fish oil, cholesterol and astaxanthin into the feed, can meet the needs of Penaeus vannamei broodstock for essential fatty acids and cholesterol, thereby increasing the egg production, hatching rate and metamorphosis rate.

[0033] 3. The environmentally friendly compound feed for Penaeus vannamei broodstock and its preparation method improve the efficacy and reduce side effects by fermenting Chinese medicine (such as astragalus, atractylodes, clove, saposhnikovia, ginsenoside, etc.) with microorganisms. The fermentation product has multiple functions: soothing the liver and regulating qi, strengthening the spleen and nourishing the stomach, enhancing flavor and fragrance, clearing away heat and detoxifying, and enhancing disease resistance and immunity. Probiotics are selected: Clostridium butyricum, yeast, Bacillus subtilis, etc. for Chinese medicine fermentation. The feed produced contains antibacterial ingredients and probiotics, which can not only inhibit harmful bacteria, but also help digestion and improve water quality, thereby improving the body's disease resistance and immunity.

[0034] The effective ingredients of traditional Chinese medicine are increased through microbial fermentation: after fermentation, the polysaccharide content in astragalus can be increased by 30%-50%, and the content of ginsenoside can be increased by 20%-30%. Enhanced pharmacological activity: the fermented traditional Chinese medicine has significantly improved antioxidant activity, immunomodulatory activity, etc., and reduced side effects.

[0035] Degradation of toxic components in traditional Chinese medicine through microbial fermentation: During the fermentation process, microorganisms can degrade the toxic components in traditional Chinese medicine and convert them into low-toxic substances.

[0036] The taste and absorption of Chinese medicine can be improved through microbial fermentation: the taste of fermented Chinese medicine is improved, it is easier to be absorbed by the human body, and the irritation to the gastrointestinal tract is reduced.

[0037] 4. The environmentally friendly formula feed for Penaeus vannamei broodstock and its preparation method add additives that promote gonad development: folic acid, soy isoflavones, krill meal, lecithin, progesterone, etc. to the feed, which promotes the rapid maturation of ovaries of broodstock, the improvement of gonad index, the large number of eggs, and the good number and quality of nauplii.

[0038] 5. The environmentally friendly compound feed for broodstock of Penaeus vannamei and the preparation method thereof can meet the nutritional requirements of broodstock breeding by adjusting the proportions of various nutritional components and selecting raw materials. The feeding effect thereof is not significantly different from that of feeding fresh lugworms in terms of spawning rate, continuous spawning rate, hatching rate, metamorphosis rate and egg diameter. Therefore, the feed of the present invention can replace fresh bait to fortify the nutrition of broodstock, and avoids the problem of infection from external pathogens due to feeding fresh bait.

[0039] Specific implementation method (blue means no modification)

[0040] The embodiments of the present invention will be described in detail below, however, the embodiments of the present invention are not limited thereto. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0041] The embodiment of the present invention provides an environmentally friendly compound feed for broodstock of Penaeus vannamei, comprising the following raw materials in weight percentage: 26.4% fish meal, 15.2% flour, 13.3% uni peptide, 12.16% fermented soybean powder, 5.7% squid powder, 3.42% shrimp slurry, 3.86% marine flavor, 2.88% fish oil, 2.66% spiral bath, 1.14% calcium dihydrogen phosphate, 0.35% salt, 0.05% astaxanthin, 0.05% cholesterol, 0.85% additive for promoting gonad development, 1.52% probiotics, 0.15% Chinese medicinal materials, 1.14% mineral trace elements, and 1.14% vitamin premix.

[0042] Among the above raw materials, the Chinese medicinal materials are formulated in mg / kg feed ratio, which includes the following raw materials in weight proportion: 5 mg of Astragalus, 6 mg of Atractylodes macrocephala, 3.5 mg of Clove, 2.5 mg of Saposhnikovia divaricata, and 12 mg of Ginsenoside; the probiotics are formulated in mg / kg feed ratio, which includes the following raw materials in weight proportion: 10 mg of Clostridium butyricum, 5 mg of yeast, and 10 mg of Bacillus subtilis; the additives for promoting gonadal development are formulated in mg / kg feed ratio, which includes the following raw materials in weight proportion: 500 mg of folic acid, 2000 mg of soy isoflavones, 800 mg of lecithin, 200 mg of progesterone, and 5000 mg of krill powder; the multivitamins are formulated in mg / kg feed ratio, which includes the following raw materials in weight proportion: Feed: Vitamin B12, 10mg, vitamin K3, 20mg, thiamine, 30mg, riboflavin, 100mg, pyridoxine hydrochloride, 30mg, inositol, 1800mg, pantothenic acid, 5430mg; niacin, 3500mg, biotin, 200mg, vitamin A, 60mg, vitamin D, 20mg, vitamin E, 200mg; complex minerals are formulated in mg / kg feed ratio, including the following raw materials in weight proportions: sodium fluoride 30mg, potassium iodide 100mg, cobalt chloride 300mg, copper sulfate 500mg, iron sulfate 400mg, zinc sulfate 250mg, magnesium sulfate 120g, calcium dihydrogen phosphate 10000mg, and zeolite powder 3500mg.

[0043] Based on the above raw material ratio, the present invention also provides a method for preparing an environmentally friendly compound feed for Penaeus vannamei broodstock, and the specific preparation steps are as follows:

[0044] S1. First, the fermented Chinese medicine is prepared. The probiotics selected are: Clostridium butyricum, yeast, and Bacillus subtilis. The Chinese medicine used is: Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata, and Ginsenosides. The solid fermentation method is used to ferment Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata, and Ginsenosides, including raw material processing, strain preparation, fermentation material preparation, inoculation and fermentation, fermentation post-processing, and fermentation effect.

[0045] S2. Prepare the raw materials according to the above weight percentages, and then mix and stir them evenly; crush the mixed raw materials through an ultrafine grinder so that all ingredients can pass through a 200-320um sieve;

[0046] S3, then move the material into the modem, pass 90-95°C steam into it for modulation for 5-10 minutes, and then extrude and granulate it at 85-90°C to form broodstock feed with a specification of 1.5×3.0mm;

[0047] S4, drying the feed pellets at 80-85°C;

[0048] S5. After the dried pellet feed is cooled, it can be packaged and stored in a cool and dark place.

[0049] The research and development principle of the present invention: It has been found that the efficacy of Chinese medicine can be improved and the side effects can be reduced after microbial fermentation. Special Chinese medicines are selected: astragalus, atractylodes, cloves, saposhnikovia and ginsenosides; astragalus has the effects of enhancing the body's immune function, protecting the liver, diuresis, anti-aging, anti-stress and lowering blood pressure; atractylodes has the effects of replenishing qi and strengthening the spleen, drying dampness and promoting diuresis, stopping sweating and stabilizing pregnancy; cloves have the effects of soothing the liver, regulating qi, strengthening the spleen and nourishing the stomach, it can treat spleen and stomach disharmony and indigestion, as well as discomfort symptoms such as loss of appetite and abdominal distension, and enhance the taste and fragrance , which is also its most important effect; the effect of windproof is to clear away heat, detoxify, dispel wind and dampness; ginsenosides have a good anti-tumor effect, can improve the body's disease resistance and immunity, and have a good effect of relieving physical strength and resisting fatigue; probiotics are selected: Clostridium butyricum, yeast, Bacillus subtilis, etc. for traditional Chinese medicine fermentation, and the feed produced contains antibacterial ingredients and probiotics, which can not only inhibit harmful bacteria, but also aid digestion and improve water quality, thereby improving the body's disease resistance and immunity.

[0050] Among them, Clostridium butyricum has a very strong intestinal regulating effect. It can inhibit pathogenic bacteria in the intestine and promote the growth of beneficial bacteria in the intestine such as bifidobacteria and lactobacillus. The use of Clostridium butyricum as a new feed additive is of great significance in reducing the abuse of antibiotic products in current feed, reducing drug residues in meat products, reducing the drug resistance of animal bacteria and ensuring animal health.

[0051] The main functions of yeast: 1. It can be used as a high-quality protein source to replace fish meal in aquatic animal feed. Yeast cells are rich in nutrients, including a variety of essential amino acids and essential fatty acids, rich vitamins, minerals, a variety of digestive enzymes and unknown growth-promoting factors. 2. Adding yeast to feed can enhance the immune function of aquatic animals. The yeast cell wall is the main part of yeast that can stimulate the body's immunity. This function of the yeast cell wall has been used in various animal breeding. 3. Yeast can grow and reproduce under both aerobic and anaerobic conditions. Adding it to water can improve water quality and increase water activity.

[0052] Bacillus subtilis has a strong inhibitory effect on harmful microorganisms such as Vibrio, Escherichia coli and baculovirus in aquaculture. It can secrete a large amount of chitinase to decompose toxic and harmful substances in the aquaculture pond and purify the water quality. At the same time, it can decompose the leftover bait, feces, organic matter, etc. in the pond, and has a strong effect of cleaning small garbage particles in the water. Bacillus subtilis is also widely used in feed. It has strong protease, lipase and amylase activities, which can promote the degradation of nutrients in feed and make aquatic animals absorb and utilize feed more fully.

[0053] The invention is based on the nutritional requirements and reproductive physiological characteristics of gonad development of broodstock of Penaeus vannamei, adopts additives capable of promoting gonad development, and develops functional broodstock feed capable of promoting gonad development. At present, broodstock feeds on the market are feeds with slightly higher nutrition than that of adult shrimps, and there is no broodstock compound feed capable of promoting gonad development in the true sense. The additives capable of promoting gonad development are mainly folic acid, soybean isoflavones, krill meal, lecithin, progesterone and the like.

[0054] In order to further better explain the present invention, the following specific embodiments are also provided:

[0055] This example aims at screening different levels of krill meal in the feed of Penaeus vannamei, designs three groups of artificial compound feeds, and carries out nutritional fortification culture of broodstock with lugworms as control. The broodstock are fed for 60 days, and their spawning performance and offspring quality are observed to prove the beneficial effects of the present invention compared with the prior art.

[0056] Specifically, the three kinds of artificial feeds for broodstock use fish meal, mussel meal, and Antarctic krill meal as protein sources and fish oil as fat sources. The formula composition and conventional nutrient content of broodstock feed are shown in Table 1. The control group is fed with sandworms. The feed raw materials are crushed and passed through a 200-320um sieve, mixed by a step-by-step enlargement method, and then stirred with water, and then pressed into a granulator to form broodstock feed with a specification of 1.5×3.0mm. After drying, store in a cool and dark place for future use.

[0057] Table 1 Test feed formula composition and nutritional composition (%)

[0058]

[0059]

[0060]

[0061]

[0062] The test water was taken from deep sea water and used after sedimentation, sand filtration and disinfection. During the nutrient fortification period, the water temperature was maintained at 27-28℃, salinity 28-33‰, pH 8.0-8.5, dissolved oxygen >5mg / L, total ammonia nitrogen <0.5mg / L, nitrite <0.1mg / L, and water depth 50-60cm.

[0063] During the fertilization and spawning period, in addition to the normal treatment, 10ppm EDTA and 0.5ppm povidone iodine solution need to be added to the spawning and hatching water to complex the heavy metal ions in the water and disinfect the water.

[0064] The experiment was divided into four treatment groups, each with 80 shrimps, with a male-female ratio of 1:1. Two parallel groups were fed with feed 1, feed 2, feed 3 and nereids (nereids were fed alive, soaked in a 50ppm potassium permanganate solution for two to three minutes before feeding and then washed with clean water). At the beginning of the experiment, an eye mark was put on one side of the eye stalk of each parent shrimp to distinguish individuals. After the end of nutritional fortification, all female parent shrimps underwent unilateral eye stalk removal surgery using tweezers. After the surgery, all treatment groups were fed with fresh nereids, and artificial insemination was used to collect reproduction and larval development data.

[0065] After artificial insemination, the egg production, hatching rate and metamorphosis rate of each treatment group were counted; the egg production rate, continuous egg production rate and average egg production number of each treatment group were calculated. About 50 fertilized eggs were taken from the first ten spawning times of each treatment group and stored in a 4% formaldehyde solution for measuring the egg diameter of the fertilized eggs; at the same time, about 0.2g of fertilized eggs were collected from the first ten spawning times of each treatment group and stored in liquid nitrogen for egg fatty acid measurement. The egg production and reproduction of parent shrimps in different treatment groups are shown in Table 2:

[0066] Table 2 Egg laying, hatching rate and metamorphosis rate of each feed group

[0067]

[0068]

[0069] Note: Different superscript letters represent significant differences (P<0.05), the same below.

[0070] As can be seen from the table, the egg laying rate of Feed Group 2 was significantly higher than that of Feed Group 1 (P<0.05), and there was no significant difference between the control group (sandworm) and Feed Group 3 (P>0.05). The control group had the largest egg laying rate of 234,500±6,300, followed by Feed Group 2, Feed Group 3 and Feed Group 1, which were 225,300±7,000, 217,400±28,600 and 180,600±6,800 respectively.

[0071] The hatching rates of the four treatment groups ranged from 45.02±1.20% to 49.26±0.82%, with the control group having the highest rate. There was no significant difference in hatching rates among the four groups (P>0.05). Among the four groups, the metamorphosis rates of the control group, feed group 2 and feed group 3 had no significant difference, but were significantly higher than those of feed group 1 (P<0.05).

[0072] The spawning rate, continuous spawning rate and average spawning times of broodstock are shown in Table 3:

[0073] Table 3 Egg laying rate, continuous egg laying rate and average egg laying times of each feed group

[0074] Feed 1 group Feed 2 Groups Feed 3 groups Nereidia group Spawning rate 70.09±1.89％ 74.16±1.18％ 69.05±3.36％ 59.17±1.18％ Continuous spawning rate 54.20±6.02％ 54.60±5.00％ 55.00±7.07％ 51.0±6.52％ Average number of spawning 1.25±0.30 1.88±0.38 2.00±0.28 1.05±0.20

[0075] Spawning rate = number of spawning female shrimps / total number of female shrimps × 100%;

[0076] Continuous spawning rate = number of female shrimps that spawn multiple times / number of female shrimps that spawn × 100%;

[0077] As can be seen from the table, the egg-laying rate of feed 2 was the highest, reaching 74.16±1.18%, the egg-laying rate of the sandworm group was the lowest, at 59.17±1.18%, the egg-laying rates of feed 1 and feed 3 were 70.09±1.89% and 69.05±3.36%, respectively, and there was no significant difference among feed 1, feed 2, and feed 3 (P>0.05); for the continuous egg-laying rate, feed 3 had the highest rate, reaching 55.00±7.07%, which was significantly higher than the control group, and there was no significant difference between feed 1 and feed 2 (P>0.05).

[0078] The average number of spawning times of each treatment group was calculated by combining all the spawning situations of each treatment group during the whole experiment. As can be seen from the table, the average number of spawning times of feed 3 group was the largest, which was 2.00±0.28, significantly higher than feed 1 and the control group, and no significant difference with feed 2 group (P>0.05).

[0079] The diameter of the eggs in the first ten spawnings of each treatment group was measured using an optical microscope. 50 eggs were laid each time, and a total of 500 fertilized eggs were tested in each treatment group. The measurement results are shown in Table 4:

[0080] Table 4 Egg diameter of each feed group

[0081] Feed 1 group Feed 2 Groups Feed 3 groups Nereidia group Egg diameter 256.70±4.54μm 263.37±3.00μm 267.84±7.19μm 58.21±8.23μm

[0082] Egg diameter = (long diameter + short diameter) / 2.

[0083] As can be seen from the table, the largest egg diameter was in feed group 3, which was 267.84±7.19μm; the smallest was in feed group 1, which was 256.70±4.54μm; feed group 2 and the control group were 263.37±3.00μm and 258.21±8.23μm, respectively. Single-factor ANOVA showed that the control group was significantly different from feed group 3 (P<0.05), and was not significantly different from feed group 1 and feed group 2 (P>0.05), and there was no significant difference between feed group 2 and feed group 3 (P>0.05).

[0084] The results of the fatty acid composition measurements of the three groups of artificial feeds and lugworms are shown in Table 5:

[0085] Table 5 Fatty acid composition of artificial compound feed and lugworm (%)

[0086] Feed 1 group Feed 2 Groups Feed 3 groups Nereidia group fatty acid 28.94±0.60％ 37.3±1.13％ 38.23±0.15％ 46.6±0.68％ n-6 series PUFA 8.43±0.01％ 8.89±0.01％ 10.41±0.01％ 13.6±0.01％ n-3 series PUFA 18.55±0.72％ 25.3±0.75％ 33.00±0.24％ 33.0±0.67％ n-3 / n-6 value 2.20±0.03 2.85±0.10 3.17±0.02 2.42±0.04

[0087] Note: The fatty acid content is expressed as the ratio of each fatty acid methyl ester to the total fatty acid methyl esters, the same below.

[0088] As can be seen from the table, the total polyunsaturated fatty acid (PUFA) content in nereid was the highest, at 46.68±0.68%; the ΣPUFA of feed 2 and feed 3 were 37.31±1.13% and 38.23±0.15% respectively, and the content of feed 1 was the lowest, only 28.94±0.60%, and there was no significant difference between feed 2 and feed 3 (P>0.05). There was no significant difference in DHA (C22:6n-3) content between the control nereid group and feed 3 (P<0.05), and the content of EPA (C20:5n-3) and AA (C20:4n-6) in nereid was higher than that of the three groups of artificial compound feeds, but in comparison, feed 2 and feed 3 had EPA and AA content closer to that of nereid, and there was no significant difference between the two groups.

[0089] The content of n-6 series PUFA was the highest in nereids, reaching 13.63±0.01%, which was significantly different from the other groups (P<0.05); the content of n-3 series PUFA in nereids was also higher than that in the other three groups of artificial feeds, reaching 33.05±0.67%. However, the n-3 / n-6 value was the largest in feed 3, which was 3.17±0.02, followed by feed 2, nereids and feed 1, with values ​​of 2.85±0.10, 2.42±0.04 and 2.20±0.03 respectively.

[0090] Fatty acid composition of fertilized eggs in each treatment group:

[0091] The ΣPUFA content in eggs of the control group was the highest, at 24.59±1.18%, followed by the feed group 2, at 23.62±0.48%, with no significant difference between the two groups (P>0.05); the ΣPUFA content in the feed group 3 was 22.93±1.01%, with no significant difference between the two groups (P>0.05), and the ΣPUFA content in the feed group 1 was the lowest, at 19.14±0.07%, with significant difference between the other three groups (P<0.05). The EPA content in eggs of the control group was the highest, at 9.16±1.03%, and the contents in the feed groups 2 and 3 were 8.37±0.02% and 8.88±0.04%, respectively, with no significant difference between the three groups (P>0.05); the EPA content in the feed group 1 was 7.60±0.38%, with significant difference between the sandworm group. The AA content in the eggs of the control group was lower than that of the feed groups 2 and 3, but higher than that of the feed group 1. The content of AA in the feed group 3 was the highest, at 1.48±0.28%. The DHA content in the eggs of the feed group 1 was the lowest, and there was a significant difference with the other three groups (P<0.05). There was no significant difference between the feed groups 2, 3 and the sandworm group (P>0.05). The n-3 / n-6 ratio in the four treatment groups ranged from 1.21±0.14 to 1.36±0.25, and there was no significant difference among the four groups (P>0.05). The feed group 3 had the highest content, at 1.36±0.25, followed by the feed group 2.

[0092] Table 6 Fatty acid composition of eggs in each treatment group (%)

[0093]

[0094]

[0095] From the above experiments, it can be seen that adding 10% krill meal by mass into the feed of Penaeus vannamei broodstock, together with 40% fish meal, 6% casein, 15% mussel powder, 14.75% high-gluten flour, 0.5% cholesterol, 2% phospholipids, 7% fish oil, 1% choline, 0.5% astaxanthin, 0.75% vitamin C, 1.5% calcium hydrogen phosphate, 0.5% multivitamins and 0.5% multiminerals, can increase the spawning rate and continuous spawning rate of Penaeus vannamei broodstock; at the same time, the hatching rate, metamorphosis rate and egg diameter are also improved to varying degrees, which can be comparable to those of sandworms.

[0096] In summary, the environmentally friendly formula feed for Penaeus vannamei broodstock and the preparation method thereof are highly resistant and multi-spawning formula feed for Penaeus vannamei broodstock. The formula feed is made by fermenting Chinese medicine with probiotics, combining with additives for promoting gonad development, screening out protein sources, amino acids, lipids, vitamins and mineral elements, and thereby ensuring that the immunity and disease resistance of the broodstock are improved by feeding the formula feed, while making the breeding water of the broodstock stable and lasting; achieving the indicators such as rapid maturation of the ovaries of the broodstock, improved gonad index, large egg carrying capacity, and good quantity and quality of nauplii.

[0097] Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that various changes, modifications, substitutions and variations may be made to the embodiments without departing from the principles and spirit of the present invention, and that the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. An environmentally friendly feed for Penaeus vannamei broodstock, characterized in that: The invention comprises the following raw materials in weight percentage: 26.4% fish meal, 15.2% flour, 13.3% uni peptide, 12.16% fermented soybean powder, 5.7% squid powder, 3.42% shrimp paste, 3.86% marine flavor, 2.88% fish oil, 2.66% spiral bath, 1.14% calcium dihydrogen phosphate, 0.35% salt, 0.05% astaxanthin, 0.05% cholesterol, 0.85% additive for promoting gonad development, 1.52% probiotics, 0.15% Chinese medicinal materials, 1.14% mineral trace elements and 1.14% vitamin premix.

2. The environmentally friendly formulated feed for Penaeus vannamei broodstock according to claim 1, characterized in that: The Chinese medicinal materials are formulated in mg / kg feed ratio, which includes the following raw materials in weight proportions: 5 mg of Astragalus, 6 mg of Atractylodes macrocephala, 3.5 mg of Clove, 2.5 mg of Saposhnikovia divaricata, and 12 mg of Ginsenoside.

3. The environmentally friendly formulated feed for Penaeus vannamei broodstock according to claim 1, characterized in that: The probiotics are formulated in mg / kg feed ratio, and include the following raw materials in weight proportions: 10 mg of Clostridium butyricum, 5 mg of yeast, and 10 mg of Bacillus subtilis.

4. The environmentally friendly formulated feed for broodstock of Penaeus vannamei according to claim 1, characterized in that: The additive for promoting gonadal development is formulated in mg / kg feed ratio, and includes the following raw materials in proportion by weight: 500 mg folic acid, 2000 mg soy isoflavones, 800 mg lecithin, 200 mg progesterone, and 5000 mg krill meal.

5. The environmentally friendly formulated feed for broodstock of Penaeus vannamei according to claim 1, characterized in that: The compound vitamin is formulated in mg / kg feed ratio, and includes the following raw materials in weight proportions: vitamin B12, 10 mg, vitamin K3, 20 mg, thiamine, 30 mg, riboflavin, 100 mg, pyridoxine hydrochloride, 30 mg, inositol, 1800 mg, pantothenic acid, 5430 mg; niacin, 3500 mg, biotin, 200 mg, vitamin A, 60 mg, vitamin D, 20 mg, and vitamin E, 200 mg.

6. The environmentally friendly formulated feed for Penaeus vannamei broodstock according to claim 1, characterized in that: The composite mineral is formulated in mg / kg feed ratio, and includes the following raw materials in weight proportions: 30 mg sodium fluoride, 100 mg potassium iodide, 300 mg cobalt chloride, 500 mg copper sulfate, 400 mg iron sulfate, 250 mg zinc sulfate, 120 g magnesium sulfate, 10000 mg calcium dihydrogen phosphate, and 3500 mg zeolite powder.

7. A method for preparing the highly resistant and multi-spawning Litopenaeus vannamei broodstock feed according to claim 1, characterized in that: S1. First, the fermented Chinese medicine is prepared. The probiotics selected are: Clostridium butyricum, yeast, and Bacillus subtilis. The Chinese medicine used is: Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata, and Ginsenosides. The solid fermentation method is used to ferment Astragalus, Atractylodes macrocephala, Clove, Saposhnikovia divaricata, and Ginsenosides, including raw material processing, strain preparation, fermentation material preparation, inoculation and fermentation, fermentation post-processing, and fermentation effect. S2. Prepare the raw materials according to the above weight percentages, and then mix and stir them evenly; crush the mixed raw materials through an ultrafine grinder so that all ingredients can pass through a 200-320um sieve; S3, then move the material into the modem, pass 90-95°C steam into it for modulation for 5-10 minutes, and then extrude and granulate it at 85-90°C to form broodstock feed with a specification of 1.5×3.0mm; S4, drying the feed pellets at 80-85°C; S5. After the dried pellet feed is cooled, it can be packaged and stored in a cool and dark place.

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