Compound microorganism aquatic feed for enhancing disease resistance of shrimps and preparation process of compound microorganism aquatic feed

Abstract

The invention relates to the technical field of aquatic feed, in particular to compound microorganism aquatic feed capable of enhancing disease resistance of shrimps and a preparation process of the compound microorganism aquatic feed. The preparation method comprises the following steps: adding honeysuckle, fructus chebulae, purslane, yucca, radix isatidis and liquorice, heating and extracting, then carrying out enzymolysis by adopting bromelain and ficin which are specifically matched, then adding peptone, glucose and specific zymophyte species, namely lactobacillus casei subsp. Casei CICC 6113 and bacillus subtillis CICC 22063, fermenting, then adding ethanol, and carrying out freeze-drying to obtain a finished product. The preparation method comprises the following steps: firstly, mixing the raw materials, adjusting the concentration of ethanol in a feed liquid, further extracting, then preparing a compound microbial fermentation product through processes such as filtration, ultrafiltration impurity removal and drying, and matching the compound microbial fermentation product with a basal feed to prepare the aquatic feed capable of effectively promoting the growth of the shrimps and efficiently improving the disease resistance of the shrimps.

Description

Technical Field

[0001] The invention relates to the technical field of aquatic feed, and in particular to a composite microbial aquatic feed capable of enhancing the disease resistance of shrimps and a preparation process thereof. Background Art

[0002] The development of my country's aquatic feed industry has generally gone through three stages. The first stage was before the 1980s, when feed was basically based on natural feed; the second stage was from the 1980s to the late 1990s, when China's aquatic feed industry began to develop, technology and market gradually took shape, and the annual output of the feed industry jumped to second place in the world; the third stage was after 2000, when industry policies became increasingly standardized, market concentration increased, innovation became the winning strategy for enterprises, and the number of feed varieties increased.

[0003] Aquatic feed for shrimp is an important category of aquatic feed. There are many kinds of raw materials for shrimp feed, mainly including animal and plant ingredients. Animal raw materials such as fish meal and silkworm pupae provide high-quality protein and essential amino acids, which are essential for the growth and development of shrimp. Plant raw materials such as corn, soybean meal and peanut cake provide rich carbohydrates and cellulose, which help to increase the energy value of feed and promote digestion. In addition, additives such as vitamins, minerals and probiotics are added to provide comprehensive nutritional support, enhance the resistance of shrimp, and improve the digestion and absorption rate of feed. The reasonable combination of these raw materials can meet the nutritional needs of shrimp at different growth stages and promote their rapid growth and high yield.

[0004] As the breeding environment changes and deteriorates, shrimp feed needs to enhance the resistance and antibacterial properties of shrimp in addition to providing nutrition. Therefore, providing a composite microbial aquatic feed that enhances the disease resistance of shrimp is a technical problem that needs to be solved at present. Summary of the invention

[0005] In view of the above technical problems, the present invention proposes a technical solution of the present invention, which includes a composite microbial aquatic feed for enhancing the disease resistance of shrimps and a preparation process thereof.

[0006] The invention provides a composite microbial aquatic feed for enhancing the disease resistance of shrimps, comprising: a composite microbial fermentation product and a balance of basic feed.

[0007] The composite microbial aquatic feed for enhancing the disease resistance of shrimps provided by the invention comprises, by mass percentage, 0.08-5% of composite microbial fermentation products and the remainder of basic feed.

[0008] Furthermore, the basic feed raw materials include: fish meal, soybean meal, kelp meal, flour, calcium lactate, vitamin E acetate, calcium dihydrogen phosphate, grape seed oil, taurine, vitamin B, glycine, and lysine. The above basic feed provides daily nutrition for shrimps, promotes the weight gain of shrimps, and also provides functions such as anti-oxidation and inhibition of pathogens.

[0009] Furthermore, the basic feed includes, by mass, 20-30 parts of fish meal, 15-30 parts of soybean meal, 5-15 parts of kelp meal, 20-30 parts of flour, 0.3-1 parts of calcium lactate powder, 0.2-1 parts of vitamin E acetate, 0.2-1 parts of calcium dihydrogen phosphate, 1-3 parts of grape seed oil, 0.05-0.3 parts of taurine, 0.02-0.3 parts of vitamin B, 0.01-0.1 parts of glycine, and 0.01-0.1 parts of lysine.

[0010] Furthermore, the composite microbial fermentation product is obtained by microbial fermentation of honeysuckle, terminalia chebula, purslane, yucca, isatis root, and licorice, which can enhance the resistance of shrimp, enhance the disease resistance of shrimp, and promote the growth of shrimp.

[0011] Furthermore, the preparation method of the composite microbial fermentation product comprises the following steps:

[0012] Step 1: crush honeysuckle, terminalia chebula, purslane, yucca, isatis root, and licorice, mix to obtain a mixed powder, add water, heat and extract to obtain an extract;

[0013] Step 2: adding bromelain and ficin to the extract for enzymolysis, inactivating the enzymes, and obtaining an enzymolysate;

[0014] Step 3: sterilizing the enzymatic hydrolysate, adding peptone and glucose to obtain a material, inoculating fermentation bacteria into the material, fermenting, and obtaining a fermented material;

[0015] Step 4: sterilize the fermented material, add ethanol, heat and stir to obtain the intermediate material;

[0016] Step 5: filtering, ultrafiltering and drying the intermediate material to obtain the composite microbial fermentation product.

[0017] Furthermore, in terms of mass parts, the amounts of raw materials used in step 1 are: 0.5-2 parts of honeysuckle, 1-3 parts of terminalia chebula, 0.5-1.5 parts of purslane, 1.5-4 parts of yucca, 0.4-1 parts of isatis root and 0.5-1 parts of licorice.

[0018] Furthermore, the mass ratio of Terminalia chebula to Yucca can be 1:1.5-3.

[0019] Furthermore, in step 1, the mass of water added is 10-30 times that of the mixed powder.

[0020] Furthermore, in step 1, the heating extraction conditions are: heating to 40-60° C. and stirring for 1-4 hours.

[0021] Furthermore, the total amount of bromelain and ficin added is 0.005-0.04 times the mass of the mixed powder, and the mass ratio of bromelain to ficin is 1:2-3.

[0022] Furthermore, in step 2, the enzymatic hydrolysis temperature is 25-45° C. and the time is 3-7 hours.

[0023] Furthermore, the amount of peptone added is 0.001-0.01 times the mass of the mixed powder.

[0024] Furthermore, the amount of glucose added is 0.01-0.045 times the mass of the mixed powder.

[0025] Furthermore, the fermentation bacteria are composed of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a bacterial count of 0.2-0.6:1.

[0026] Furthermore, the amount of fermentation bacteria in the material is 3.3×10 10 to 6.8×10 10 cfu / mL.

[0027] Furthermore, in step 3, the fermentation temperature is 30-45° C. and the fermentation time is 50-100 hours.

[0028] Furthermore, the mass of ethanol added in step 4 is 0.25-0.45 times the mass of water added in step 1.

[0029] Furthermore, in step 4, the heating and stirring conditions are heating to 40-55° C. for 2-6 hours.

[0030] Furthermore, in step 5, the ultrafiltration is performed using an ultrafiltration membrane with a molecular weight cut-off lower than 5 KDa.

[0031] The present invention also provides a method for preparing the composite microbial aquatic feed for enhancing the disease resistance of shrimps, comprising the steps of mixing, granulating and sterilizing the raw materials.

[0032] Beneficial effects of the present invention:

[0033] The invention adopts honeysuckle, terminalia chebula, purslane, yucca, isatis root and liquorice to add and heat for extraction, then adopts specially matched bromelain and ficin for enzymolysis, then adds peptone and glucose, and specially matched fermentation bacteria species Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 for fermentation, then adds ethanol, adjusts the ethanol concentration in the feed liquid, further extracts, and then prepares a composite microbial fermentation product through filtering, ultrafiltration to remove impurities, drying and other processes, and matches the composite microbial fermentation product with a basic feed to prepare an aquatic feed that effectively promotes shrimp growth and can efficiently improve the disease resistance of shrimp.

[0034] The feed prepared by the invention can effectively increase the weight of shrimps, give shrimps better growth effects, and can also effectively increase the resistance of shrimps to pathogens and increase the survival rate of shrimps infected with viruses.

[0035] The invention adopts bromelain and ficin enzymatic hydrolysis treatment to improve the growth effect and disease resistance of shrimp to a certain extent. The two are enzymatically hydrolyzed together to increase the types and concentrations of nutritional and antibacterial active ingredients, thereby improving the feeding and functional effects of the feed.

[0036] The present invention adopts composite bacteria for fermentation, which can greatly improve the effect of enhancing shrimp resistance in feed, so that shrimp has good growth effect and excellent anti-bacteria effect. In particular, when Lactobacillus paracasei subspecies CI CC6113 and Bacillus subtilis CI CC 22063 are used in specific amounts, the antibacterial effect of shrimp can be synergistically improved, and when the ratio of the fermentation bacteria of the two is 0.2-0.6:1, the active ingredients produced by fermentation have better effects, and the feed has better performance.

[0037] The invention adds ethanol and adjusts the ethanol concentration, which can enhance the anti-bacterial effect of the composite microbial fermentation product and effectively improve the growth performance of shrimp. The combination of terminalia chebula and yucca is conducive to improving the feed and increasing the resistance of shrimp. DETAILED DESCRIPTION

[0038] The present invention will be described below in conjunction with specific embodiments, and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific embodiments are used to illustrate the present invention, rather than to limit the present invention.

[0039] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0040] Bromelain: 100,000 U / g, Guangdong Mingtong Biotechnology Co., Ltd.

[0041] Fig proteinase: 100,000 U / g, Nanjing Tongying Biotechnology Co., Ltd.

[0042] Lactobacillus casei subsp. casei CICC 6113: Code: CICC 6113, commercially purchased from China Industrial Microbiological Culture Collection Administration Center.

[0043] Bacillus subtilis CICC 22063: No. CICC 22063, commercially purchased from China Industrial Microbiological Culture Collection Administration Center.

[0044] Peptone: Product number: P304956, soybean peptone, Shanghai Aladdin Biochemical Technology Co., Ltd.

[0045] Preparation Example 1:

[0046] The method for preparing the composite microbial fermentation product comprises the following steps:

[0047] Step 1: Grind the dried honeysuckle buds, terminalia chebula fruits, purslane leaves, yucca roots, isatis roots, and licorice roots to a particle size of less than 0.1 mm, mix to obtain a mixed powder, add water 20 times the mass of the mixed powder, heat to 50° C., stir at 75 rpm for 3 hours for heating extraction, and obtain an extract;

[0048] Step 2: add bromelain and ficin to the extract, perform enzymolysis at 38°C for 4.5 hours, inactivate the enzymes, and obtain an enzymolysate;

[0049] Step 3: sterilize the enzymatic hydrolysate, add 0.005 times the mass of peptone and 0.032 times the mass of glucose of the mixed powder to obtain a material, inoculate fermentation bacteria into the material, and ferment at 37.2° C. for 87 hours to obtain a fermented material;

[0050] Step 4: sterilize the fermentation material, add ethanol, heat to 50°C, stir at 80rpm for 5.3 hours to obtain an intermediate material;

[0051] Step 5: The intermediate material is filtered using a filter membrane with a pore size of 2 microns, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1.2 KDa to obtain an ultrafiltrate, and the ultrafiltrate is freeze-dried to a water content of 2.93% to obtain the composite microbial fermentation product.

[0052] The fermentation bacteria consisted of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a ratio of 0.5:1.

[0053] The amount of fermentation bacteria in the material is 5.5×10 10 cfu / mL;

[0054] The raw materials used in step 1 are as follows: 0.8 parts of honeysuckle, 1.5 parts of terminalia chebula, 0.7 parts of purslane, 3 parts of yucca, 0.7 parts of isatis root and 0.8 parts of liquorice;

[0055] The mass ratio of terminalia chebula to yucca is 1:2;

[0056] The total amount of bromelain and ficin added is 0.025 times the mass of the mixed powder, and the mass ratio of bromelain to ficin is 1:2.5;

[0057] The mass of ethanol added in step 4 is 0.4 times the mass of water added in step 1.

[0058] Preparation Example 2:

[0059] The method for preparing the composite microbial fermentation product comprises the following steps:

[0060] Step 1: Grind the dried honeysuckle buds, terminalia chebula fruits, purslane leaves, yucca roots, isatis roots, and licorice roots to a particle size of less than 0.1 mm, mix to obtain a mixed powder, add water 22 times the mass of the mixed powder, heat to 57° C., stir at 75 rpm for 2.5 hours for heating extraction, and obtain an extract;

[0061] Step 2: adding bromelain and ficin to the extract, performing enzymolysis at 37°C for 5 hours, inactivating the enzymes, and obtaining an enzymolysate;

[0062] Step 3: sterilize the enzymatic hydrolysate, add 0.005 times the mass of peptone and 0.032 times the mass of glucose of the mixed powder to obtain a material, inoculate fermentation bacteria into the material, and ferment at 37.5° C. for 85 hours to obtain a fermented material;

[0063] Step 4: sterilize the fermentation material, add ethanol, heat to 52°C, and stir at 80 rpm for 5.5 hours to obtain an intermediate material;

[0064] Step 5: The intermediate material is filtered using a filter membrane with a pore size of 2 microns, and the filtrate is ultrafiltered using an ultrafiltration membrane with a molecular weight cutoff of 1.2 KDa to obtain an ultrafiltrate, and the ultrafiltrate is freeze-dried to a water content of 2.89% to obtain the composite microbial fermentation product.

[0065] The fermentation bacteria consisted of Lactobacillus casei subspecies CI CC 6113 and Bacillus subtilis CI CC 22063 in a bacterial number of 0.35:1;

[0066] The amount of fermentation bacteria in the material is 5.7×10 10 cfu / mL;

[0067] The raw materials used in step 1 are as follows: 0.5 parts of honeysuckle, 1.3 parts of terminalia chebula, 1 part of purslane, 3.2 parts of yucca, 0.8 parts of isatis root and 0.7 parts of liquorice;

[0068] The mass ratio of terminalia chebula to yucca is 1:3;

[0069] The total amount of bromelain and ficin added was 0.026 times the mass of the mixed powder, and the mass ratio of bromelain to ficin was 1:2.46;

[0070] The mass of ethanol added in step 4 is 0.38 times the mass of water added in step 1.

[0071] Preparation Example 3:

[0072] The only difference from Preparation Example 1 is that step 2 is: the extract is placed at 38° C. for 4.5 hours to obtain a treated product. The rest is the same as Preparation Example 1.

[0073] Preparation Example 4:

[0074] The only difference from Preparation Example 1 is that: Step 2 is: add bromelain in an amount of 0.025 times the mass of the mixed powder to the extract, perform enzymolysis at 38° C. for 4.5 hours, inactivate the enzyme, and obtain an enzymolyzate. The rest is the same as Preparation Example 1.

[0075] Preparation Example 5:

[0076] The only difference compared with Preparation Example 1 is that: Step 2 is: adding 0.025 times the mass of the mixed powder to the extract, performing enzymolysis at 38° C. for 4.5 hours, inactivating the enzyme, and obtaining an enzymolyzate. The rest is the same as Preparation Example 1.

[0077] Preparation Example 6:

[0078] The only difference compared with Preparation Example 1 is that: Step 3 is: sterilize the enzymolyzate, add 0.005 times the mass of peptone and 0.032 times the mass of glucose of the mixed powder to obtain a material, and place it at 37.2° C. for 87 hours to obtain a material. The rest is the same as Preparation Example 1.

[0079] Preparation Example 7:

[0080] The only difference compared with Preparation Example 1 is that: Step 3 is: sterilize the enzymolyzate, add 0.005 times the mass of the mixed powder peptone and 0.032 times the mass of glucose to obtain a material, inoculate the material with fermentation bacteria Lactobacillus casei subspecies CICC 6113, and the amount of fermentation bacteria in the material is 5.5×10 10 cfu / mL, and fermented at 37.2°C for 87 hours to obtain a fermented material. Other details were the same as in Preparation Example 1.

[0081] Preparation Example 8:

[0082] The only difference compared with Preparation Example 1 is that: Step 3 is: sterilize the enzymatic hydrolysate, add 0.005 times the mass of peptone and 0.032 times the mass of glucose of the mixed powder to obtain a material, inoculate the material with fermentation bacteria Bacillus subtilis CICC 22063, and the amount of fermentation bacteria in the material is 5.5×10 10 cfu / mL, and fermented at 37.2°C for 87 hours to obtain a fermented material. Other details were the same as in Preparation Example 1.

[0083] Preparation Example 9:

[0084] The only difference compared to Preparation Example 1 is that in step 3, the fermentation bacteria are composed of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a ratio of 0.05:1, and the amount of the fermentation bacteria in the material is 5.5×10 10 cfu / mL. Others are the same as in Preparation Example 1.

[0085] Preparation Example 10:

[0086] The only difference compared with Preparation Example 1 is that in step 3, the fermentation bacteria are composed of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a ratio of 1:1, and the amount of the fermentation bacteria in the material is 5.5×10 10 cfu / mL. Others are the same as in Preparation Example 1.

[0087] Preparation Example 11:

[0088] The only difference compared with Preparation Example 1 is that the mass of ethanol added in step 4 is 0.65 times the mass of water added in step 1. The rest is the same as Preparation Example 1.

[0089] Preparation Example 12:

[0090] The only difference compared with Preparation Example 1 is that the mass of ethanol added in step 4 is 0.1 times the mass of water added in step 1. The rest is the same as Preparation Example 1.

[0091] Preparation Example 13:

[0092] The raw material dosage in step 1 is: 0.8 parts of honeysuckle, 4.5 parts of terminalia chebula, 0.7 parts of purslane, 0.7 parts of isatis root and 0.8 parts of liquorice. Other ingredients are the same as those in Preparation Example 1.

[0093] Preparation Example 14:

[0094] The raw material dosage in step 1 is: 0.8 parts of honeysuckle, 0.7 parts of purslane, 4.5 parts of yucca, 0.7 parts of isatis root and 0.8 parts of liquorice. Other ingredients are the same as those in Preparation Example 1.

[0095] The composite microbial fermentation product prepared in the above preparation example is used to prepare a composite microbial aquatic feed that enhances shrimp disease resistance. The details are as follows:

[0096] 1. Feed 1

[0097] Raw material composition, wt%: 0.25% of the composite microbial fermentation product of Preparation Example 1 and the remainder of the basic feed.

[0098] Among them, the basic feed composition is: 22.5 parts of fish meal, 20.3 parts of soybean meal, 8.9 parts of kelp meal, 24.3 parts of flour, 0.6 parts of calcium lactate powder, 0.35 parts of vitamin E acetate, 0.3 parts of calcium dihydrogen phosphate, 1.5 parts of grape seed oil, 0.1 parts of taurine, 0.05 parts of vitamin B, 0.03 parts of glycine, and 0.03 parts of lysine.

[0099] 2. Feed 2

[0100] Raw material composition, wt%: 0.25% of the composite microbial fermentation product of Preparation Example 2 and the remainder of the basic feed.

[0101] Among them, the basic feed composition is: 23.7 parts of fish meal, 18.5 parts of soybean meal, 10.7 parts of kelp meal, 23.4 parts of flour, 0.7 parts of calcium lactate powder, 0.38 parts of vitamin E acetate, 0.35 parts of calcium dihydrogen phosphate, 1.25 parts of grape seed oil, 0.09 parts of taurine, 0.055 parts of vitamin B, 0.032 parts of glycine, and 0.028 parts of lysine.

[0102] 3. Feed 3

[0103] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 3 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0104] 4. Feed 4

[0105] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 4 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0106] Feed 5

[0107] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 5 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0108] Feed 6

[0109] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 6 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0110] Feed 7

[0111] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 7 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0112] Feed 8

[0113] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 8 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0114] Feed 9

[0115] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 9 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0116] Feed 10

[0117] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 10 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0118] Feed 11

[0119] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 11 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0120] Feed 12

[0121] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 12 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0122] Feed 13:

[0123] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 13 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0124] Feed 14:

[0125] The difference from Feed 1 is that the composite microbial fermentation product of Preparation Example 14 is used to replace the composite microbial fermentation product of Preparation Example 1, and the rest is the same as Feed 1.

[0126] The preparation method of the above feed is as follows: various raw materials are mixed, granulated and sterilized to obtain the feed.

[0127] The above feed was used to feed shrimps and the effect was tested.

[0128] The specific method is as follows:

[0129] Raw material preparation: the above feeds 1-14 and blank group feed. The blank group feed composition is: 18.2 parts of fish meal, 17.3 parts of soybean meal, 25.3 parts of flour, 0.27 parts of calcium lactate powder, 0.4 parts of calcium dihydrogen phosphate, 1.6 parts of grape seed oil, 0.12 parts of taurine, 0.07 parts of vitamin B, and 0.03 parts of glycine.

[0130] Take the same batch of white shrimp cultured, and use the above control feed for adaptive feeding for 5 days, feed at 5% of body weight, and feed twice a day. After the adaptive feeding is completed, white shrimp of similar size are grouped into 15 groups, namely feed groups 1-14 and blank control group, with 200 tails in each group. Each group is fed at 5% of body weight, twice a day, among which feed groups 1-14 are fed with feed 1-14 in turn, and the blank control group is fed with blank group feed; feed for 20 consecutive days, during which the water temperature is controlled at 22-26℃ and oxygenated. Before feeding, weigh the total weight of the shrimp and calculate the average weight of each shrimp; after 20 days of feeding, test the number of deaths of white shrimp in each group, weigh the total weight of live shrimp and calculate the average weight, and then calculate the weight gain rate. The results are shown in Table 1.

[0131]

[0132]

[0133] After the above-mentioned 20-day feeding, 130 white shrimps in each group were randomly selected for disease resistance testing. Specifically, 130 shrimps were selected from both feed groups 1-14 and the blank control group and fed with shrimp meat infected with white spot virus for one attack. Then, feeding was carried out: feed groups 1-14 were fed with their respective corresponding feeds, and the blank control group was fed with blank group feed; the feeding was continued for 7 days, during which the water temperature was controlled at 22-26℃ and oxygenated. On the 4th and 7th days of feeding, the cumulative number of deaths of white shrimps in each group was observed, and the cumulative mortality rate was calculated. The results are shown in Table 2.

[0134] Test / Cumulative number of deaths (only) Day 4 Day 7 Blank control group 98 130 Feed 1 group 44 73 Feed 2 Groups 45 75 Feed 3 groups 57 91 Feed 4 groups 48 79 Feed 5 groups 46 77 Feed 6 groups 65 102 Feed 7 groups 54 87 Feed 8 groups 53 86 Feed 9 groups 48 78 Feed 10 groups 50 81 Feed 11 groups 51 83 Feed 12 groups 55 88 Feed 13 groups 47 77 Feed 14 groups 48 79

[0135] The cumulative number of deaths in Table 2 was converted into the cumulative mortality rate, and the results are shown in Table 3.

[0136]

[0137]

[0138] The test results in Tables 1-3 show that the feed prepared by the present invention can effectively increase the weight of shrimps, give shrimps better growth effects, and can effectively increase the resistance of shrimps to pathogens and increase the survival rate of shrimps infected with viruses.

[0139] The test results of feeds 1, 3-5 show that the enzymatic hydrolysis of bromelain and ficin in the present invention improves the growth effect and disease resistance of shrimp to a certain extent. The two enzymes are hydrolyzed together to increase the types and concentrations of nutritional and antibacterial active ingredients, thereby improving the feeding and functional effects of the feed.

[0140] It can be seen from the test results of feeds 1, 6-10 that the present invention uses composite bacteria for fermentation, which can greatly improve the effect of feed on enhancing shrimp resistance, so that shrimp has a good growth effect and excellent anti-bacteria effect. In particular, when Lactobacillus paracasei subspecies casei CICC 6113 and Bacillus subtilis CICC 22063 are used in specific amounts, the antibacterial effect of shrimp can be synergistically improved, and when the ratio of the fermented bacteria of the two is 0.2-0.6:1, the active ingredients produced by fermentation have a better effect, and the feed has better performance.

[0141] The test results of feeds 1, 11-12 show that the present invention adds ethanol and adjusts the ethanol concentration to enhance the anti-bacterial effect of the composite microbial fermentation product and effectively improve the growth performance of shrimp. The combination of terminalia chebula and yucca is beneficial to improve the resistance of shrimp in feed. The above content is obtained in combination with the test results of feeds 1, 13-14.

[0142] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A composite microbial aquatic feed for enhancing the disease resistance of shrimp, characterized in that: The content of the compound microbial fermentation product and the balance of the basic feed is as follows: The preparation method of the composite microbial fermentation product comprises the following steps: Step 1: crush honeysuckle, terminalia chebula, purslane, yucca, isatis root, and licorice, mix to obtain a mixed powder, add water, heat and extract to obtain an extract; Step 2: adding bromelain and ficin to the extract for enzymolysis, inactivating the enzymes, and obtaining an enzymolysate; Step 3: sterilizing the enzymatic hydrolysate, adding peptone and glucose to obtain a material, inoculating fermentation bacteria into the material, fermenting, and obtaining a fermented material; Step 4: sterilize the fermented material, add ethanol, heat and stir to obtain the intermediate material; Step 5: filtering, ultrafiltering and drying the intermediate material to obtain the composite microbial fermentation product.

2. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: The basic feed includes, by mass, 20-30 parts of fish meal, 15-30 parts of soybean meal, 5-15 parts of kelp meal, 20-30 parts of flour, 0.3-1 parts of calcium lactate powder, 0.2-1 parts of vitamin E acetate, 0.2-1 parts of calcium dihydrogen phosphate, 1-3 parts of grape seed oil, 0.05-0.3 parts of taurine, 0.02-0.3 parts of vitamin B, 0.01-0.1 parts of glycine, and 0.01-0.1 parts of lysine.

3. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: The fermentation bacteria are composed of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a bacterial count of 0.2-0.6:

1.

4. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: The amount of fermentation bacteria in the material is 3.3×10 10 to 6.8×10 10 cfu / mL.

5. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: In terms of mass parts, the amounts of raw materials used in step 1 are: 0.5-2 parts of honeysuckle, 1-3 parts of terminalia chebula, 0.5-1.5 parts of purslane, 1.5-4 parts of yucca, 0.4-1 parts of isatis root and 0.5-1 parts of licorice.

6. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: The mass ratio of Terminalia chebula to Yucca is 1:1.5-3.

7. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: In step 1, the mass of water added is 10-30 times that of the mixed powder; and / or, the total amount of bromelain and ficin added is 0.005-0.04 times the mass of the mixed powder, and the mass ratio of bromelain to ficin is 1:2-3; And / or, the amount of peptone added is 0.001-0.01 times the mass of the mixed powder; and / or, the amount of glucose added is 0.01-0.045 times the mass of the mixed powder; And / or, the mass of ethanol added in step 4 is 0.25-0.45 times the mass of water added in step 1.

8. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: In step 1, the heating extraction conditions are: heating to 40-60°C and stirring for 1-4 hours; And / or, in step 2, the enzymatic hydrolysis temperature is 25-45°C and the time is 3-7 hours; and / or, in step 3, the fermentation temperature is 30-45° C. and the fermentation time is 50-100 hours; And / or, in step 4, the heating and stirring conditions are heating to 40-55° C. for 2-6 hours.

9. The composite microbial aquatic feed for enhancing disease resistance of shrimp according to claim 1, characterized in that: In step 5, the ultrafiltration is performed using an ultrafiltration membrane with a molecular weight cut-off lower than 5 KDa.

10. A method for preparing a composite microbial aquatic feed for enhancing shrimp disease resistance according to any one of claims 1 to 9, characterized in that: The process includes mixing, granulating and sterilizing the raw materials.

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