A compound microbial aquatic feed for enhancing the disease resistance of shrimps and its preparation process

Abstract

The present invention relates to the technical field of aquatic feeds, and particularly relates to a compound microbial aquatic feed for enhancing the disease resistance of shrimps and its preparation process. In the present invention, honeysuckle, Terminalia chebula, Portulaca oleracea, Yucca schidigera, Isatis indigotica, and liquorice are added and heated for extraction, then enzymatically hydrolyzed with specifically paired bromelain and ficin, and then peptone and glucose are added, as well as specific fermentation bacteria species Lactobacillus casei subsp. casei CICC 6113 and Bacillus subtilis CICC 22063 for fermentation. Then ethanol is added to adjust the ethanol concentration in the feed liquid for further extraction, and then a compound microbial fermentation product is prepared through processes such as filtration, ultrafiltration for impurity removal, and drying. The compound microbial fermentation product is combined with a basic feed to prepare an aquatic feed that can effectively promote the growth of shrimps and highly enhance the disease resistance of shrimps.

Description

Technical Field

[0001] The present invention relates to the technical field of aquatic feeds, and particularly to a compound microbial aquatic feed for enhancing the disease resistance of shrimps and its preparation process. Background Art

[0002] The development of China's aquatic feed industry has generally gone through three stages. The first stage was before the 1980s, when the feeds were basically natural feeds. The second stage was from the 1980s to the end of the 1990s. The Chinese aquatic feed industry began to develop, technologies and markets gradually took shape, and the annual output of the feed industry ranked second in the world. The third stage was after 2000. Industry policies became increasingly standardized, the degree of market centralization increased, innovation became the key to winning for enterprises, and the types of feeds gradually increased.

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

[0004] With the change and deterioration of the aquaculture environment, in addition to providing nutrition, shrimp feeds also need to enhance the resistance and antibacterial properties of shrimps. Therefore, providing a compound microbial aquatic feed for enhancing the disease resistance of shrimps is a technical problem that needs to be solved currently. Summary of the Invention

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

[0006] The present invention provides a compound microbial aquatic feed for enhancing the disease resistance of shrimps, including: a compound microbial fermentation product and the remaining basic feed.

[0007] The compound microbial aquatic feed for enhancing the disease resistance of shrimps provided by the present invention includes, by mass percentage: 0.08 - 5% of the compound microbial fermentation product and the remaining basic feed.

[0008] Furthermore, the basic feed ingredients include: fish meal, soybean meal powder, kelp powder, flour, calcium lactate, vitamin E acetate, calcium dihydrogen phosphate, grape seed oil, taurine, vitamin B, glycine, and lysine. The above basic feed provides the daily nutrition of shrimps, promotes the weight gain of shrimps, and also provides functions such as antioxidant and antibacterial effects.

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

[0010] Furthermore, the compound microbial fermentation product is obtained by microbial fermentation of honeysuckle, terminalia chebula, portulaca oleracea, yucca, isatis root, and licorice, which can enhance the resistance of shrimps, improve their disease resistance, and promote their growth.

[0011] Furthermore, the preparation method of the compound microbial fermentation product includes the following steps:

[0012] Step 1: Honeysuckle, terminalia chebula, portulaca oleracea, yucca, isatis root, and licorice are crushed and mixed to obtain a mixed powder. Water is added and heated for extraction to obtain an extract.

[0013] Step 2: Bromelain and ficin are added to the extract for enzymatic hydrolysis, and then the enzymes are inactivated to obtain an enzymolysate.

[0014] Step 3: The enzymolysate is sterilized, peptone and glucose are added to obtain a material, and fermentation bacteria are inoculated into the material for fermentation to obtain a fermented material.

[0015] Step 4: The fermented material is sterilized, ethanol is added, and the mixture is heated and stirred to obtain an intermediate material.

[0016] Step 5: The intermediate material is filtered, ultrafiltered, and dried to obtain the compound microbial fermentation product.

[0017] Furthermore, by mass, the raw material dosage in Step 1 is as follows: 0.5 - 2 parts of honeysuckle, 1 - 3 parts of terminalia chebula, 0.5 - 1.5 parts of portulaca oleracea, 1.5 - 4 parts of yucca, 0.4 - 1 part of isatis root, and 0.5 - 1 part 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] Further, in step 1, the heating extraction conditions are: heating to 40 - 60°C and stirring for 1 - 4 hours.

[0021] Further, 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] Further, in step 2, the enzymatic hydrolysis temperature is 25 - 45°C and the time is 3 - 7 hours.

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

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

[0025] Further, the fermenting bacteria are composed of Lactobacillus paracasei subsp. paracasei CICC 6113 and Bacillus subtilis CICC 22063 with a bacterial quantity ratio of 0.2 - 0.6:1.

[0026] Further, the dosage of the fermenting bacteria in the material is 3.3×10 10 to 6.8×10 10 cfu / mL.

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

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

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

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

[0031] The present invention also provides a preparation method of the compound microbial aquatic feed for enhancing the disease resistance of shrimps, including the steps of mixing, granulating and sterilizing each raw material.

[0032] Advantages of the present invention:

[0033] The present invention uses honeysuckle, Terminalia chebula, purslane, yucca, isatis root, and licorice, adds them and heats for extraction, then uses bromelain and ficin with a specific combination for enzymatic hydrolysis, adds peptone and glucose, and specific fermentation bacteria types Lactobacillus casei subsp. casei CICC 6113 and Bacillus subtilis CICC 22063 for fermentation, then adds ethanol, adjusts the ethanol concentration in the feed liquid for further extraction, and then prepares a composite microbial fermentation product through processes such as filtration, ultrafiltration for impurity removal, and drying, and matches the composite microbial fermentation product with a basic feed to prepare an aquatic feed that can effectively promote the growth of shrimp and highly improve the disease resistance ability of shrimp.

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

[0035] The present invention uses bromelain and ficin for enzymatic hydrolysis, which can improve the growth effects and disease resistance of shrimp to a certain extent. The two are enzymatically hydrolyzed together to increase the types and concentrations of nutrients and antibacterial active ingredients, and improve the feeding and functional effects of the feed.

[0036] The present invention uses composite bacteria for fermentation, which can greatly improve the effect of the feed in enhancing the resistance of shrimp, making shrimp have better growth effects and excellent efficacy in resisting pathogens. Especially when using Lactobacillus casei subsp. casei CI CC6113 and Bacillus subtilis CI CC 22063 with specific dosage combinations, it can synergistically enhance the antibacterial effect of shrimp, and when the ratio of the two fermentation bacteria is 0.2 - 0.6:1, the active ingredients produced by fermentation play a better effect, and the feed has better performance.

[0037] The present invention adds ethanol and adjusts the ethanol concentration, which can enhance the effect of the composite microbial fermentation product in resisting pathogens and effectively improve the growth performance of shrimp. The combination of Terminalia chebula and yucca is beneficial to improving the resistance of shrimp by the feed. Detailed implementation methods

[0038] The following will elaborate on the present invention in combination with specific implementation methods, and various effects of the present invention will be presented more clearly therefrom. Those skilled in the art should understand that these specific implementation methods are used to illustrate the present invention rather than limit the present invention.

[0039] Unless otherwise specifically stated, various raw materials, reagents, instruments, and equipment used in the present invention can be obtained through market purchase or can be prepared by existing methods.

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

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

[0042] Lactobacillus casei subsp. casei CICC 6113: Number: CICC 6113, commercially purchased from China Center of Industrial Culture Collection.

[0043] Bacillus subtilis CICC 22063: Number CICC 22063, commercially purchased from China Center of Industrial Culture Collection.

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

[0045] Preparation Example 1:

[0046] The preparation method of the composite microbial fermentation product comprises the following steps:

[0047] Step 1: Dry Flos Lonicerae buds, Fructus Chebulae, Portulaca oleracea L. leaves, Yucca root, Isatis root, and Glycyrrhiza uralensis Fisch. roots are pulverized to a particle size below 0.1 mm, mixed to obtain a mixed powder, 20 times the mass of water of the mixed powder is added, heated to 50 °C, and stirred at 75 rpm for 3 hours for heat extraction to obtain an extract;

[0048] Step 2: Bromelain and ficin are added to the extract, and enzymatic hydrolysis is carried out at a temperature of 38 °C for 4.5 hours, and the enzyme is inactivated to obtain an enzymolysate;

[0049] Step 3: The enzymolysate is sterilized, 0.005 times the mass of peptone and 0.032 times the mass of glucose of the mixed powder are added to obtain a material, fermentation bacteria are inoculated into the material, and fermentation is carried out at a temperature of 37.2 °C for 87 hours to obtain a fermentation material;

[0050] Step 4: The fermentation material is sterilized, ethanol is added, heated to 50 °C, and stirred at 80 rpm for 5.3 hours to obtain an intermediate material;

[0051] Step 5: The intermediate material is filtered with a filter membrane with a pore size of 2 μm, the filtrate is ultrafiltered with an ultrafiltration membrane with a molecular weight cut-off of 1.2 kDa, the ultrafiltrate is freeze-dried to a water content of 2.93% by mass to obtain the composite microbial fermentation product.

[0052] Among them, the fermentation bacteria are composed of Lactobacillus casei subsp. casei CICC 6113 and Bacillus subtilis CICC 22063 with a bacterial quantity ratio of 0.5:1;

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

[0054] In Step 1, the amounts of raw materials used are: 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 licorice root;

[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] In Step 4, the mass of ethanol added is 0.4 times the mass of water added in Step 1.

[0058] Preparation Example 2:

[0059] The preparation method of the composite microbial fermentation product comprises the following steps:

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

[0061] Step 2: Add bromelain and ficin to the extract, carry out enzymatic hydrolysis at 37 °C for 5 hours, inactivate the enzyme to obtain an enzymolyzate;

[0062] Step 3: Sterilize the enzymolyzate, add 0.005 times the mass of the mixed powder of peptone and 0.032 times the mass of glucose to obtain a material, inoculate fermentation bacteria in the material, ferment at 37.5 °C for 85 hours to obtain a fermentation material;

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

[0064] Step 5: Filter the intermediate material with a filter membrane with a pore size of 2 microns, ultrafilter the filtrate with an ultrafiltration membrane with a cut-off molecular weight of 1.2 kDa, obtain an ultrafiltrate, and lyophilize the ultrafiltrate to a water content of 2.89% by mass to obtain the composite microbial fermentation product.

[0065] Among them, the fermentation bacteria are composed of Lactobacillus paracasei subsp. paracasei CICC 6113 and Bacillus subtilis CICC 22063 with a bacterial quantity ratio of 0.35:1;

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

[0067] In Step 1, the dosages of raw materials are as follows: 0.5 part of honeysuckle, 1.3 parts of Terminalia chebula Retz., 1 part of Portulaca oleracea L., 3.2 parts of Yucca schidigera, 0.8 part of Isatis indigotica Fortune, and 0.7 part of liquorice;

[0068] The mass ratio of Terminalia chebula Retz. to Yucca schidigera is 1:3;

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

[0070] In Step 4, the mass of ethanol added is 0.38 times the mass of water added in Step 1.

[0071] Preparation Example 3:

[0072] The difference compared with Preparation Example 1 is only that: in Step 2, the extract is left standing at a temperature of 38°C for 4.5 hours to obtain a treated product. Others are the same as Preparation Example 1.

[0073] Preparation Example 4:

[0074] The difference compared with Preparation Example 1 is only that: in Step 2, 0.025 times the mass of the mixed powder of bromelain is added to the extract, and enzymatic hydrolysis is carried out at a temperature of 38°C for 4.5 hours, and then the enzyme is inactivated to obtain an enzymolyzed product. Others are the same as Preparation Example 1.

[0075] Preparation Example 5:

[0076] The difference compared with Preparation Example 1 is only that: in Step 2, 0.025 times the mass of the mixed powder of ficin is added to the extract, and enzymatic hydrolysis is carried out at a temperature of 38°C for 4.5 hours, and then the enzyme is inactivated to obtain an enzymolyzed product. Others are the same as Preparation Example 1.

[0077] Preparation Example 6:

[0078] The difference compared with Preparation Example 1 is only that: in Step 3, the enzymolyzed product is sterilized, 0.005 times the mass of the mixed powder of peptone and 0.032 times the mass of glucose are added to obtain a material, and the material is placed at a temperature of 37.2°C for 87 hours to obtain a material. Others are the same as Preparation Example 1.

[0079] Preparation Example 7:

[0080] The difference compared with Preparation Example 1 is only that: in Step 3, the enzymolyzed product is sterilized, 0.005 times the mass of the mixed powder of peptone and 0.032 times the mass of glucose are added to obtain a material, and the fermentation fungus Lactobacillus casei subsp. casei CICC 6113 is inoculated into the material, and the dosage of the fermentation fungus in the material is 5.5×10 10 cfu / mL, and fermentation is carried out at a temperature of 37.2°C for 87 hours to obtain a fermented material. Others are the same as Preparation Example 1.

[0081] Preparation Example 8:

[0082] The difference compared with Preparation Example 1 is only that: in Step 3, the enzymolysate is sterilized, peptone 0.005 times the mass of the mixed powder and glucose 0.032 times the mass of the mixed powder are added to obtain a material, and the fermentation bacterium Bacillus subtilis CICC 22063 is inoculated into the material. The dosage of the fermentation bacterium in the material is 5.5×10 10 cfu / mL, and fermentation is carried out at 37.2 °C for 87 hours to obtain a fermented material. Others are the same as Preparation Example 1.

[0083] Preparation Example 9:

[0084] The difference compared with Preparation Example 1 is only that: in Step 3, the fermentation bacterium consists of Lactobacillus paracasei subsp. paracasei CICC 6113 and Bacillus subtilis CICC 22063 with a bacterial quantity ratio of 0.05:1, and the dosage of the fermentation bacterium in the material is 5.5×10 10 cfu / mL. Others are the same as Preparation Example 1.

[0085] Preparation Example 10:

[0086] The difference compared with Preparation Example 1 is only that: in Step 3, the fermentation bacterium consists of Lactobacillus paracasei subsp. paracasei CICC 6113 and Bacillus subtilis CICC 22063 with a bacterial quantity ratio of 1:1, and the dosage of the fermentation bacterium in the material is 5.5×10 10 cfu / mL. Others are the same as Preparation Example 1.

[0087] Preparation Example 11:

[0088] The difference compared with Preparation Example 1 is only that: in Step 4, the mass of ethanol added is 0.65 times the mass of water added in Step 1. Others are the same as Preparation Example 1.

[0089] Preparation Example 12:

[0090] The difference compared with Preparation Example 1 is only that: in Step 4, the mass of ethanol added is 0.1 times the mass of water added in Step 1. Others are the same as Preparation Example 1.

[0091] Preparation Example 13:

[0092] The amounts of raw materials used in Step 1 are: 0.8 parts of honeysuckle, 4.5 parts of Terminalia chebula, 0.7 parts of Portulaca oleracea, 0.7 parts of Isatis tinctoria, and 0.8 parts of liquorice. Others are the same as Preparation Example 1.

[0093] Preparation Example 14:

[0094] In Step 1, the amounts of raw materials used are as follows: 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 licorice. Others are the same as in Preparation Example 1.

[0095] Use the composite microbial fermentation product prepared in the above preparation example to prepare a composite microbial aquatic feed for enhancing the disease resistance of shrimps. Specifically 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 balance of the basal feed.

[0098] Among them, the basal feed composition is: 22.5 parts of fish meal, 20.3 parts of soybean meal powder, 8.9 parts of kelp powder, 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, 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 balance of the basal feed.

[0101] Among them, the basal feed composition is: 23.7 parts of fish meal, 18.5 parts of soybean meal powder, 10.7 parts of kelp powder, 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, 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 others are 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 others are 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 others are the same as Feed 1.

[0108] Feed 6

[0109] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 6 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0110] Feed 7

[0111] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 7 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0112] Feed 8

[0113] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 8 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0114] Feed 9

[0115] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 9 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0116] Feed 10

[0117] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 10 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0118] Feed 11

[0119] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 11 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0120] Feed 12

[0121] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 12 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0122] Feed 13:

[0123] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 13 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0124] Feed 14:

[0125] It is different from Feed 1 in that the compound microbial fermentation product of Preparation Example 14 is used to replace the compound microbial fermentation product of Preparation Example 1, and the others are the same as Feed 1.

[0126] The preparation method of the above feeds is as follows: Mix, granulate and sterilize each raw material to obtain the feed.

[0127] Feed the prawns with the above feed and conduct an effect test.

[0128] The specific method is as follows:

[0129] Raw material preparation: The above feeds 1-14 and the blank group feed. Among them, the composition of the blank group feed is: 18.2 parts of fish meal, 17.3 parts of soybean meal powder, 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] Select Litopenaeus vannamei cultured in the same batch, and adaptively feed them with the above control feed for 5 days, feeding at 5% of the body weight, twice a day. After the adaptive feeding, select Litopenaeus vannamei with similar sizes for grouping. Set 15 groups, namely feeds 1-14 groups and the blank control group, with 200 tails in each group. Each group is fed at 5% of the body weight, twice a day. Among them, the feeds 1-14 groups are fed with feeds 1-14 in sequence, and the blank control group is fed with the blank group feed; continuously feed for 20 days. During the feeding period, the water temperature is controlled at 22-26 °C, and oxygenation treatment is carried out. Before feeding, weigh the total weight of the shrimps and calculate the average weight of each shrimp; after 20 days of feeding, test the death number of Litopenaeus vannamei in each group and weigh the total weight of the live shrimps 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 20 days of feeding, randomly select 130 Litopenaeus vannamei from each group for disease resistance testing. Specifically: 130 shrimps are selected from the feeds 1-14 groups and the blank control group and fed with the shrimp meat infected with white spot virus for one challenge treatment. Then carry out feeding: the feeds 1-14 groups are fed with their corresponding feeds, and the blank control group is fed with the blank group feed; continuously feed for 7 days. During the feeding period, the water temperature is controlled at 22-26 °C, and oxygenation treatment is carried out. Observe the cumulative death number of Litopenaeus vannamei in each group on the 4th and 7th days of feeding, and calculate the cumulative mortality rate. The results are shown in Table 2.

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

[0135] Convert the cumulative death number in Table 2 into the cumulative mortality rate, and the results are shown in Table 3.

[0136]

[0137]

[0138] From the test results in Table 1-3, it can be seen that the feed prepared by the present invention can effectively increase the weight of shrimp, endow the shrimp with good growth effects, and at the same time effectively increase the resistance of shrimp to pathogens and increase the survival rate of shrimp infected with viruses.

[0139] From the test results of Feed 1, 3-5, it can be seen that the present invention uses bromelain and ficin for enzymatic hydrolysis, which to a certain extent improves the growth effects and disease resistance of shrimp. The two are enzymatically hydrolyzed together to increase the types and concentrations of nutrients and antibacterial active ingredients, thus enhancing the feeding and functional effects of the feed.

[0140] From the test results of Feed 1, 6-10, it can be seen that the present invention uses compound bacteria for fermentation, which can greatly improve the effect of the feed in enhancing the resistance of shrimp, enabling the shrimp to have good growth effects and excellent efficacy in resisting pathogens. Especially when specific dosages of Lactobacillus casei subsp. casei CICC 6113 and Bacillus subtilis CICC 22063 are combined, the antibacterial effect of shrimp can be synergistically enhanced. When the ratio of the two fermenting bacteria is 0.2-0.6:1, the active ingredients produced by fermentation exert better effects and the feed has better performance.

[0141] From the test results of Feed 1, 11-12, it can be obtained that the present invention adds ethanol and adjusts the ethanol concentration, which can enhance the effect of the compound microbial fermentation product in resisting pathogens and effectively improve the growth performance of shrimp. The combination of Terminalia chebula and Yucca schidigera is beneficial to improving the resistance of the feed to shrimp. The above content is obtained by combining the test results of Feed 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 equivalent technologies, the present invention is also intended to include these changes and modifications.

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; In terms of weight, the raw materials used in step 1 are as follows: 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 liquorice; the weight ratio of terminalia chebula to yucca is 1:1.5-3; The fermentation bacteria consisted of Lactobacillus casei subspecies CICC 6113 and Bacillus subtilis CICC 22063 in a ratio of 0.2-0.6:1; the amount of the fermentation bacteria in the material was 3.3×10 10 to 6.8×10 10 cfu / mL; 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.

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: In step 1, the mass of water added is 10-30 times that of the mixed powder; 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.

4. 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.

5. 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.

6. A method for preparing a composite microbial aquatic feed for enhancing shrimp disease resistance according to any one of claims 1 to 5, characterized in that: The process includes mixing, granulating and sterilizing the raw materials.

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