阅读说明：本技术 含有酵母细胞壁的异育银鲫配合饲料及其应用 (Carassius auratus gibelio compound feed containing yeast cell walls and application thereof ) 是由 易建华 俞学锋 李知洪 胡骏鹏 杨凡 聂琴 于 2018-08-30 设计创作，主要内容包括：本发明涉及一种含有酵母细胞壁的异育银鲫配合饲料,其含有白鱼粉、植物蛋白源、鱼油、植物油、氯化胆碱和酵母细胞壁。本发明的异育银鲫配合饲料可以改善异育银鲫生长性能,同时增强异育银鲫机体免疫力,提高抗病能力。通过该饲料在异育银鲫养殖中的应用,可以实现异育银鲫无公害健康养殖技术,达到替代传统化学药物的使用的目的,减少药物残留和环境污染。(The invention relates to a carassius auratus gibelio compound feed containing yeast cell walls, which contains white fish meal, a vegetable protein source, fish oil, vegetable oil, choline chloride and yeast cell walls. The carassius auratus gibelio compound feed can improve the growth performance of carassius auratus gibelio, enhance the immunity of carassius auratus gibelio bodies and improve the disease resistance. By applying the feed to carassius auratus gibelio breeding, a pollution-free healthy carassius auratus gibelio breeding technology can be realized, the purpose of replacing the traditional chemical medicine is achieved, and the medicine residue and the environmental pollution are reduced.)

1. The carassius auratus gibelio compound feed is characterized by comprising white fish meal, a vegetable protein source, fish oil, vegetable oil, choline chloride and yeast cell walls.

2. The crucian carp compound feed according to claim 1, wherein the compound feed comprises one or more than two of wheat bran, flour, corn starch, vitamin premix and mineral premix, preferably the compound feed comprises one or more than two of 4-6 wt% of wheat bran, 4-6 wt% of flour, 4-6 wt% of corn starch, 0.3-0.4 wt% of vitamin premix and 4-6 wt% of mineral premix, and further preferably the compound feed comprises 4-6 wt% of wheat bran, 4-6 wt% of flour, 4-6 wt% of corn starch, 0.3-0.4 wt% of vitamin premix and 4-6 wt% of mineral premix.

3. Crucian carp compound feed according to claim 1 or 2, wherein the compound feed comprises 9-12 wt% white fish meal, 53-66 wt% vegetable protein source, 1.5-4.5 wt% fish oil, 1.5-4.5 wt% vegetable oil, 0.05-0.2 wt% choline chloride and 0.1-1.4 wt% yeast cell wall, preferably the compound feed comprises 0.33-1.0 wt% yeast cell wall, more preferably the compound feed comprises 0.67-1.0 wt% yeast cell wall.

4. The carassius auratus gibelio compound feed as claimed in any one of claims 1-3, wherein the vegetable protein source is selected from one or more of soybean, black soybean, pea, broad bean, bean pulp, bean cake, soybean meal, cotton meal, rapeseed meal, corn protein powder, corn germ meal, peanut meal, flax meal and sesame cake; preferably, the vegetable protein source is selected from one or more than two of bean pulp, cotton pulp and rapeseed pulp; it is further preferred that the vegetable protein source comprises soybean meal, cottonseed meal, and rapeseed meal.

5. The carassius auratus gibelio compound feed as claimed in claim 4, wherein the mass percentage of the soybean meal in the compound feed is 28-32%; the mass percentage of the cottonseed meal in the compound feed is 9-14%; the mass percentage of the rapeseed meal in the compound feed is 15-20%.

6. A carassius auratus gibelio compound feed as claimed in any one of claims 1-5, wherein the vegetable oil is selected from one or more of soybean oil, linseed oil, rapeseed oil, olive oil, palm oil, corn oil and coconut oil; preferably, the vegetable oil is selected from one or more of soybean oil, rapeseed oil and coconut oil; it is further preferred that the vegetable oil is soybean oil.

7. A carassius auratus gibelio compound feed as claimed in any one of claims 1-6, wherein the compound feed comprises 9-12 wt% of white fish meal, 28-32 wt% of soybean meal, 15-20 wt% of rapeseed meal, 9-14 wt% of cottonseed meal, 4-6 wt% of flour, 4-6 wt% of corn starch, 1.5-4.5 wt% of fish oil, 1.5-4.5 wt% of soybean oil, 0.3-0.4 wt% of vitamin premix, 4-6 wt% of mineral premix, 0.05-0.2 wt% of choline chloride, 2-4 wt% of carboxymethyl cellulose and 0.1-1.4 wt% of yeast cell wall.

8. The crucian carp compound feed according to claim 7, wherein the compound feed comprises 10-12 wt% of white fish meal, 28-32 wt% of soybean meal, 15-18 wt% of rapeseed meal, 12-14 wt% of cottonseed meal, 4-6 wt% of flour, 4-5 wt% of corn starch, 3-4 wt% of fish oil, 3-4 wt% of soybean oil, 0.3-0.4 wt% of vitamin premix, 4-6 wt% of mineral premix, 0.05-0.2 wt% of choline chloride, 2-4 wt% of carboxymethyl cellulose and 0.67-1.0 wt% of yeast cell wall.

9. A carassius auratus gibelio compound feed as claimed in any one of claims 1-8, wherein the protein content in the white fish meal is above 60 wt%, preferably 65-68 wt%.

10. A carassius auratus gibelio compound feed as claimed in any one of claims 1-9, wherein the yeast cell wall contains more than 18 wt% of β -glucan and more than 17 wt% of mannooligosaccharide, preferably, the yeast cell wall contains 20-30 wt% of β -glucan and 20-35 wt% of mannooligosaccharide.

11. The crucian carp compound feed as claimed in any of claims 1-10, wherein the mass ratio of the fish oil to the vegetable oil is 3: 1-1: 3, preferably 2: 1-1: 2.

12. A method for preparing the allogynogenetic crucian carp compound feed as claimed in any one of claims 1 to 11, which is obtained by mixing raw materials containing white fish meal, a vegetable protein source, fish oil, vegetable oil, choline chloride and yeast cell walls.

13. The use of the crucian carp compound feed of any one of claims 1-11 in feeding crucian carp or improving the immunity of crucian carp.

Technical Field

The invention relates to a fish feed, in particular to an allogynogenetic crucian carp compound feed containing yeast cell walls, and belongs to the field of feed processing.

Background

Carassius auratus gibelio is a new Carassius auratus breeding object successfully developed by fish breeding experts of aquatic organism research institute of Chinese academy of sciences in 1976-1981, and is a filial generation bred through artificial insemination by using natural gynogenesis square Carassius auratus as a female parent and using Xingguo red carp as a male parent. Due to the rapid growth and short breeding period, the breeding method has wide breeding nationwide at present. However, with the increasing degree of the culture intensification, diseases of carassius auratus gibelio frequently occur. Taking the example of crucian carp bred in Jiangsu salt city, the red gill appears in 2009, the large-scale outbreak starts in 2011, other crucian carps in the same pond do not attack the disease, and the attack crucian carps can attack the disease from water bloom to adult crucian carps. The disease onset time starts from the end of 3 months, the disease onset is reduced after 11 months, and sporadic death occurs in winter. The death rate of the disease is high, and once the disease is treated improperly, the death of the whole pool is possible.

Sunqi et al (case analysis of the disease and the hemorrhagic disease of gill in the county of Kyoho, Jiangsu and she Yang, scientific fish culture, 2012(11):64-65) consider that the disease and the hemorrhagic disease of gill should be clearly distinguished, and research finds that the disease of gill with large red is bacterial infection and can occur in crucian with any specification, and the hemorrhagic disease of gill is viral infection and mainly occurs at about 0.4 kg/tail (which is going to go out of a pond), and for the disease of gill with large red, authors consider that advance prevention is needed to prevent the disease of gill with large red from being caused subsequently. In the aquaculture production, the effect of preventing and treating bacterial diseases of cultured fishes by antibiotics is increasingly poor due to the generation of drug resistance of pathogenic bacteria. Therefore, enhancing the functions of the autoimmune system of the cultured fishes and improving the disease resistance of the cultured fishes becomes an important way for researchers at home and abroad to discuss the disease prevention and treatment method of the aquatic animals at present.

The yeast cell wall is a product obtained by concentrating and drying cell walls obtained by performing autolysis or exogenous enzyme catalytic hydrolysis or mechanical crushing on thalli obtained by fermenting saccharomyces cerevisiae, and the product is rich in β -glucan, mannan oligosaccharide and other immune enhancement factors, and can enhance the immunity of organisms and improve the disease resistance.

Hu soldiers and the like (research on adding effect of yeast cell walls in carassius auratus gibelio feed, Chinese feed, 2007(20):28-30.) research on application effect of carassius auratus gibelio feed by adding yeast cell walls. The result shows that the addition of 0.10% of yeast cell wall in the feed has obvious promotion effect on the nonspecific immunity of carassius auratus gibelio and has certain promotion effect on the development of RNA/DNA and intestinal tract in the dorsal muscle of the carassius auratus gibelio.

Disclosure of Invention

The technical problem existing in the prior art is that carassius auratus gibelio is easy to be infected by bacteria in the feeding process, for example, diseases such as gill of red branchia and the like are caused by infection of aeromonas hydrophila, and the effect of preventing and treating bacterial diseases of cultured fishes by means of antibiotics is increasingly poor due to the generation of drug resistance of pathogenic bacteria.

In the prior art, 0.10% of yeast cell wall is added into the feed to obviously promote the nonspecific immunity of the carassius auratus gibelio, but the gill disease of the red crucian can occur in carassius auratus of any specification, the bacterial resistance of carassius auratus gibelio is weaker than that of adult carassius auratus gibelio, and the immunoprotection of the carassius auratus gibelio is required to be further improved.

The invention aims to solve the technical problems and finds that the growth performance of the carassius auratus gibelio can be improved and the immunity of the carassius auratus gibelio can be effectively enhanced by feeding the carassius auratus gibelio with the compound feed with a specific formula containing components such as yeast cell walls and the like in the growth process of the carassius auratus gibelio.

Specifically, the invention provides the following technical scheme:

the invention provides a carassius auratus gibelio compound feed, which contains white fish meal, a vegetable protein source, fish oil, vegetable oil, choline chloride and yeast cell walls.

Preferably, the mixed feed for carassius auratus gibelio comprises one or more than two of wheat bran, flour, corn starch, vitamin premix and mineral premix, preferably, the mixed feed comprises one or more than two of 4-6 wt% of wheat bran, 4-6 wt% of flour, 4-6 wt% of corn starch, 0.3-0.4 wt% of vitamin premix and 4-6 wt% of mineral premix, and further preferably, the mixed feed comprises 4-6 wt% of wheat bran, 4-6 wt% of flour, 4-6 wt% of corn starch, 0.3-0.4 wt% of vitamin premix and 4-6 wt% of mineral premix.

Preferably, the above mentioned carassius auratus gibelio compound feed contains 9-12 wt% of white fish meal, 53-66 wt% of vegetable protein source, 1.5-4.5 wt% of fish oil, 1.5-4.5 wt% of vegetable oil, 0.05-0.2 wt% of choline chloride and 0.1-1.4 wt% of yeast cell wall, preferably, the compound feed contains 0.33-1.0 wt% of yeast cell wall, and more preferably, the compound feed contains 0.67-1.0 wt% of yeast cell wall.

Preferably, the above mentioned carassius auratus gibelio compound feed, wherein the vegetable protein source is selected from one or more of soybean, black soybean, pea, broad bean, bean pulp, bean cake, soybean meal, cotton pulp, rapeseed meal, corn protein powder, corn germ meal, peanut meal, flax meal and sesame cake; preferably, the vegetable protein source is selected from one or more than two of bean pulp, cotton pulp and rapeseed pulp; it is further preferred that the vegetable protein source comprises soybean meal, cottonseed meal, and rapeseed meal.

Preferably, the carassius auratus gibelio compound feed is characterized in that the soybean meal accounts for 28-32% of the mass of the compound feed; the mass percentage of the cottonseed meal in the compound feed is 9-14%; the mass percentage of the rapeseed meal in the compound feed is 15-20%.

Preferably, the above mixed feed for carassius auratus gibelio is prepared by mixing vegetable oil selected from one or more of soybean oil, linseed oil, rapeseed oil, olive oil, palm oil, corn oil and coconut oil; preferably, the vegetable oil is selected from one or more of soybean oil, rapeseed oil and coconut oil; it is further preferred that the vegetable oil is soybean oil.

Preferably, the mixed feed for carassius auratus gibelio comprises 9-12 wt% of white fish meal, 28-32 wt% of soybean meal, 15-20 wt% of rapeseed meal, 9-14 wt% of cottonseed meal, 4-6 wt% of flour, 4-6 wt% of corn starch, 1.5-4.5 wt% of fish oil, 1.5-4.5 wt% of soybean oil, 0.3-0.4 wt% of vitamin premix, 4-6 wt% of mineral premix, 0.05-0.2 wt% of choline chloride, 2-4 wt% of carboxymethyl cellulose and 0.1-1.4 wt% of yeast cell wall.

Preferably, the carassius auratus gibelio compound feed comprises 10-12 wt% of white fish meal, 28-32 wt% of soybean meal, 15-18 wt% of rapeseed meal, 12-14 wt% of cottonseed meal, 4-6 wt% of flour, 4-5 wt% of corn starch, 3-4 wt% of fish oil, 3-4 wt% of soybean oil, 0.3-0.4 wt% of vitamin premix, 4-6 wt% of mineral premix, 0.05-0.2 wt% of choline chloride, 2-4 wt% of carboxymethyl cellulose and 0.67-1.0 wt% of yeast cell wall.

Preferably, the carassius auratus gibelio compound feed has a protein content of more than 60 wt% in the white fish meal, and preferably, the protein content of the white fish meal is 65 wt% -68 wt%.

Preferably, the carassius auratus gibelio compound feed comprises the yeast cell wall containing more than 18 wt% of β -glucan and more than 17 wt% of mannooligosaccharide, and preferably, the yeast cell wall contains 20-30 wt% of β -glucan and 20-35 wt% of mannooligosaccharide.

Preferably, the carassius auratus gibelio compound feed is prepared from the fish oil and the vegetable oil in a mass ratio of 3: 1-1: 3, preferably 2: 1-1: 2.

In another aspect, the invention provides a preparation method of the carassius auratus gibelio compound feed, which is obtained by mixing raw materials containing white fish meal, a vegetable protein source, fish oil, vegetable oil, choline chloride and yeast cell walls.

On the other hand, the invention also provides application of the carassius auratus gibelio compound feed in feeding carassius auratus gibelio or improving the immunity of the carassius auratus gibelio.

The beneficial effects of the invention include:

the feed of the invention is added with yeast cell walls, and simultaneously contains fish oil, vegetable oil and fish meal, which can enhance the organism immunity of the carassius auratus gibelio, promote the carassius auratus gibelio to grow rapidly and improve the disease resistance.

The present invention and its advantageous technical effects are described in detail below with reference to specific embodiments.

Detailed Description

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. All publications and patents specifically mentioned herein are incorporated herein by reference in their entirety for all purposes including describing and disclosing the chemicals, instruments, statistical analyses and methods reported in the publications that might be relevant to the invention. All references cited in this specification are to be considered as indicative of the level of skill in the art.

Fish meal (fish meal) is a high-protein powder obtained by using one or more kinds of fishes as production raw materials and performing a series of processing such as cooking, squeezing, dewatering and crushing. Fish meal is classified into white fish meal and red fish meal (brown fish meal) according to the fish used as the raw material, the white fish meal is fish meal prepared from cold water fish such as flatfish, cod, pollock and the like, and the fish meal prepared from the fish meal is called white fish meal because the fish meat color is light and the fish meal prepared from the fish meal is light. The crude protein content of the white fish meal is generally more than 60 percent. The red fish meal, also called brown fish meal, is fish meal processed by using small trash fishes such as sardine, silver carp, anchovy, tilapia, eel and the like as raw materials and leftovers of fish and shrimp food, and is named red fish meal and also called brown fish meal because the muscle of the fish is red brown and the color of the processed fish meal is dark. The crude protein content of the red fish meal is generally over 60 percent, and the content is very small compared with that of the white fish meal. The red fish meal has higher yield than white fish meal and lower price. However, the red fish meal has high content of unsaturated fatty acid and histamine, so the red fish meal is easy to oxidize and deteriorate, animals can be poisoned after taking a large amount of histamine, and the fish meal can generate a large amount of low molecular compounds and peroxides, such as aldehyde, ketone and the like after the fat is oxidized. These substances can lead to a reduction in the nutritional value of the feed and the appetite of the animal, resulting in slow growth.

The plant protein feed source comprises: soybean, black soybean, pea, broad bean, soybean meal, soybean cake, soybean meal, cottonseed cake, rapeseed meal, rapeseed cake, corn gluten meal, corn germ meal, peanut kernel cake (meal), linseed cake (meal), sunflower kernel cake (meal), sesame cake, and the like. The soybean cake (meal) is the most common plant protein feed source due to high yield, rich nutritional value and low price.

The soybean meal is a byproduct obtained by squeezing and extracting oil from full-fat soybeans, has the protein content of 42-48 percent, and contains rich essential amino acid.

The peanut kernel cake (meal) is a byproduct of peanut oil extraction by pressing peanuts. The peanut kernel cake (meal) has the protein content of 48.68 percent, is similar to the soybean meal, but is lack of methionine, lysine and threonine, has more than 14 percent of essential amino acid, contains 32.5 percent of polysaccharide, 5.61 percent of ash, has the vitamin content of 0.871mg/100g, and contains various trace elements such as magnesium, potassium, iron, calcium, selenium, sodium, phosphorus, copper and the like.

Cottonseed meal (cake) is a by-product of cottonseed after extraction of oil by a pre-pressing extraction method or a screw pressing method. The cottonseed cake (dregs) is the by-product of cottonseed after shelling and oil extraction, and the cottonseed dregs or cottonseed cake are collectively called cottonseed dregs. The cottonseed meal contains 34% -44% protein, and its protein content is similar to that of soybean cake.

The deoiled rapeseed has by-product rapeseed dregs (cake) containing 34-38% of protein, rich lysine and methionine, but only a small amount of arginine, and rich trace elements such as phosphorus, calcium, selenium, etc., especially the highest content of selenium in vegetable protein feed sources. Rapeseed meal (cake) also contains anti-nutritional factors such as protease inhibitor, isothiocyanate, thioglucoside (GS) and tannin, but people can still use the rapeseed meal (cake) as a protein source of aquatic feeds after removing the anti-nutritional factors.

Choline is an important nutrient substance for maintaining normal growth of fishes, and plays an important role in the aspects of fish neural development, liver fat transportation, organism anabolism and the like. The feed product is choline chloride, commonly called choline chloride, and the aqueous solution is viscous liquid. A powdery product containing 50% choline chloride can be prepared by adding 70% choline chloride aqueous solution into excipient dispersant such as wheat bran, rice hull powder or silicon dioxide. The equivalent amount of choline chloride is only 87% of choline, calculated on the basis of the molecular weight of the pure product.

Corn starch is also called maize starch, and is white and light yellowish powder. Soaking corn in 0.3% sulfurous acid, crushing, sieving, precipitating, drying, grinding, etc. The common product contains a small amount of fat, protein, pigment (zeaxanthin, cryptoxanthin and the like), and the like, the corn starch added into the feed can not only be bonded with the feed, but also be used as an energy substance to save the protein, and the contained pigment can improve the body color of the carassius auratus gibelio to a certain extent.

The lipid has important effect on fish growth, is an important energy substance of fish bodies, is a main source of Essential Fatty Acid (EFA), and mainly comprises fish oil, soybean oil, peanut oil, rapeseed oil, palm oil, sunflower seed oil, cottonseed oil and the like which are commonly used in aquatic animal feed. The fatty acid compositions of different fat sources have larger difference, and the growth and the feed utilization of the carassius auratus gibelio are obviously influenced.

The soybean oil contains a large amount of linoleic acid, and is probably a good fat source for carassius auratus gibelio. The main component of fish oil is triglyceride, and also contains a small amount of lipoid, phosphoglycerol ether, liposoluble vitamins and protein degradation product, etc., wherein the phospholipid mainly comprises phosphatidylcholine and phosphatidylethanolamine. The fish oil is rich in omega-3 polyunsaturated fatty acids (DHA and EPA), contains Highly Unsaturated Fatty Acids (HUFA) such as EPA and DHA, is well utilized by fish, is a main fat source of traditional aquatic feed, but the high-content HUFA in the fish oil is easily oxidized to cause the increase of the oxidation degree of the feed. The use of fish oil and soybean oil in combination significantly improved growth compared to the corresponding single fat source. This ensures the HUFA supply to promote rapid fish growth and also reduces the oxidation of the feed. When the ratio of the fish oil to the soybean oil is in the range of 1: 3-3: 1, the growth promoting effect on the carassius auratus gibelio is the best.

Wheat bran, i.e., wheat bran, a byproduct of wheat processing flour, is rich in cellulose and vitamins. The flour is a powdery substance ground by wheat and used as a sugar source to provide energy for carassius auratus gibelio.

As described above, the present invention aims to provide a carassius auratus gibelio feed containing yeast cell walls, wherein the yeast cell walls can not only enhance the immunity of carassius auratus gibelio, but also reduce the incidence of diseases.

The invention discloses a preferable carassius auratus gibelio compound feed containing yeast cell walls, which comprises the following components in parts by mass: 9-12 parts of white fish meal, 28-32 parts of soybean meal, 15-20 parts of rapeseed meal, 9-14 parts of cottonseed meal, 4-6 parts of flour, 4-6 parts of corn starch, 3-4 parts of fish oil, 3-4 parts of soybean oil, 0.3-0.4 part of vitamin premix, 4-6 parts of mineral premix, 0.05-0.2 part of choline chloride, 2-4 parts of carboxymethyl cellulose and 0.1-1.4 parts of yeast cell wall.

The invention further preferably relates to a carassius auratus gibelio compound feed containing yeast cell walls, which comprises 10 parts of white fish meal, 30 parts of soybean meal, 18 parts of rapeseed meal, 12 parts of cottonseed meal, 4.67 parts of flour, 5 parts of wheat bran, 5 parts of corn starch, 3.3 parts of fish oil, 3.2 parts of soybean oil, 0.39 part of vitamin premix, 5 parts of mineral premix, 0.11 part of choline chloride, 3 parts of carboxymethyl cellulose and 0.33 part of yeast cell walls.

The invention further preferably relates to a carassius auratus gibelio compound feed containing yeast cell walls, which comprises 10 parts of white fish meal, 30 parts of soybean meal, 18 parts of rapeseed meal, 12 parts of cottonseed meal, 4.33 parts of flour, 5 parts of wheat bran, 5 parts of corn starch, 3.3 parts of fish oil, 3.2 parts of soybean oil, 0.39 part of vitamin premix, 5 parts of mineral premix, 0.11 part of choline chloride, 3 parts of carboxymethyl cellulose and 0.67 part of yeast cell walls.

The invention further preferably relates to a carassius auratus gibelio compound feed containing yeast cell walls, which comprises 10 parts of white fish meal, 30 parts of soybean meal, 18 parts of rapeseed meal, 12 parts of cottonseed meal, 4 parts of flour, 5 parts of wheat bran, 5 parts of corn starch, 3.3 parts of fish oil, 3.2 parts of soybean oil, 0.39 part of vitamin premix, 5 parts of mineral premix, 0.11 part of choline chloride, 3 parts of carboxymethyl cellulose and 1.0 part of yeast cell walls.

The invention further preferably relates to a carassius auratus gibelio compound feed containing yeast cell walls, which comprises 10 parts of white fish meal, 30 parts of soybean meal, 18 parts of rapeseed meal, 12 parts of cottonseed meal, 3.67 parts of flour, 5 parts of wheat bran, 5 parts of corn starch, 3.3 parts of fish oil, 3.2 parts of soybean oil, 0.39 part of vitamin premix, 5 parts of mineral premix, 0.11 part of choline chloride, 3 parts of carboxymethyl cellulose and 1.33 parts of yeast cell walls.

Preferably, in the above mixed feed for carassius auratus gibelio, the protein content in the white fish meal is more than 65%, and the choline chloride content is more than 60%.

Preferably, in the carassius auratus gibelio compound feed, the yeast cell wall is prepared from a saccharomyces cerevisiae strain, the content of the main functional component β -glucan is 20-30%, and the content of mannan oligosaccharide is more than or equal to 20%.

For better understanding of the present invention, the following embodiments are further described, but the present invention is not limited to the specific embodiments. Unless otherwise stated, the processes referred to in the present invention are conventional and the starting materials referred to are commercially available. The reagents and instrumentation used in the following examples were from the following sources:

TABLE 1 reagents and apparatus used in the examples

The self-preparation method of the raw materials used in the following examples is as follows:

the preparation method of the 0.067mol/L phosphate buffer solution comprises the following steps: weighing potassium dihydrogen phosphate (KH)₂PO₄Tianjin, Kemiou chemical reagent Co., Ltd.) was dissolved in distilled water, poured into a 1000ml volumetric flask, and diluted to the scale (1000 ml).

The preparation method of the FKC bacterins comprises the following steps: inactivating aeromonas hydrophila by using a formaldehyde solution for 72 hours at the temperature of 4 ℃ to prepare an inactivated bacterial liquid. And (3) sucking the inactivated bacterial liquid by using a pipette gun, putting the inactivated bacterial liquid into a sterilized centrifugal tube, centrifuging for 5min at 10,000rpm, pouring out supernatant, collecting thalli, washing the thalli for 4-5 times by using sterile physiological saline, washing off residual formaldehyde solution in the thalli, and finally suspending the thalli in the sterile physiological saline to obtain the aeromonas hydrophila AH1 whole-bacterium inactivated vaccine.

The preparation method of the F-SA comprises the following steps: adding formalin with the final concentration of 0.3% into staphylococcus aureus liquid, shaking uniformly, placing in a constant-temperature water bath kettle at 35 ℃ for heat preservation for 24 hours, sampling and inoculating in a plate for culture, and after proving that thalli are inactivated, centrifugally cleaning for 3 times by using sterilized normal saline to obtain the F-SA.

The preparation method of the viable bacteria liquid of the aeromonas hydrophila comprises the following steps: the strain source laboratory is characterized in that the strain to be preserved is firstly inoculated in a brain heart infusion culture medium, cultured at 25 ℃ and pre-cultured for 24 hours, then is inoculated in a fresh water agar culture medium, cultured for 48 hours, collected by a centrifugal method, washed for 3 times by sterilized normal saline, diluted to a certain concentration and refrigerated for preservation.

The preparation method of the mouse anti-aeromonas hydrophila strain serum comprises the following steps: the injection dosage is determined according to the measuring result of the aeromonas hydrophila, the mice are injected with the abdominal cavity for 3 times, the interval of each time is 10 days, and the control group is injected with normal saline. About 20 μ L of blood was drawn after the third injection 3d, and the titer of anti-AH 1 serum was determined by slide agglutination 7d after the third injection, orbital bleeding and mice sacrificed; and simultaneously taking the blood of a control group to obtain negative control serum (named as control serum), placing a centrifugal tube filled with the blood obliquely to form a maximum inclined plane, placing the centrifugal tube into a refrigerator at 4 ℃ for standing overnight to naturally separate out the serum, centrifuging the serum at 4,000rpm for 15min, taking supernatant to obtain anti-AH 1 serum and the control serum, and placing the serum and the control serum into an ultra-low temperature refrigerator at-80 ℃ for storage.

The preparation method of the low-content yeast cell wall comprises the steps of adopting a saccharomyces cerevisiae strain, taking molasses as a carbon source, urea as a nitrogen source, obtaining yeast after the fermentation period is 16 hours, carrying out autolysis treatment on the yeast, controlling the salt concentration to be 1.8%, the pH value to be 5.5, the temperature to be 55 ℃ and carrying out autolysis for 15 hours, and carrying out centrifugal separation to obtain the yeast cell wall with low polysaccharide content, wherein the β -glucan content is 18.4 wt% and the mannan oligosaccharide content is 17.5 wt%.