CN108157673B - Preparation method of fermented prawn compound feed - Google Patents

Abstract

The invention discloses a preparation method of fermented shrimp compound feed. The present invention adopts the process of mixed fermentation of solid and liquid, preferably produces abundant enzymes and can co-grow Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, and carries out joint fermentation, and through aerobic and anaerobic two-step fermentation, fully utilize Aspergillus niger and Bacillus subtilis has the advantages of abundant enzymes, so that the basic fermentation raw materials can be completely decomposed, macromolecular proteins are decomposed into peptides and amino acids, cellulose is decomposed into fermentable sugars, starch is decomposed into fermentable glucose, and xylan is decomposed into fermentable glucose. Xylose, glucan, etc., so that the growth and reproduction of Lactobacillus reuteri in the anaerobic stage can be supported by a large amount of nutrients. The biological peptide content, L-lactic acid content, the number of colonies and other indicators can reflect that the growth and metabolism of microorganisms in the aerobic stage and the anaerobic stage in the preparation method proposed by the present invention are very vigorous.

Description

A kind of preparation method of fermented shrimp compound feed

Technical field:

The invention belongs to the technical field of aquatic feed and bioengineering, and particularly relates to a preparation method of a fermented shrimp compound feed.

Background technique:

my country's shrimp farming industry has formed a complete industrial system. With the continuous expansion of farming density and scale, various diseases frequently occur. Bacterial diseases caused by Vibrio sp. At present, it has seriously restricted the development of shrimp farming in my country and even in the world. Farmers usually use antibiotics in large quantities. Although they have achieved results in a short period of time, they have led to problems such as increased resistance of pathogenic microorganisms, decreased diversity of microbial communities, drug residues, and deterioration of the breeding environment. The preparation method of the invention uses several functional strains such as Aspergillus niger, Bacillus, lactic acid bacteria, yeast, etc., through anaerobic fermentation and then aerobic fermentation of shrimp compound feed, pre-expanding or crushing of feed materials, etc., and through microbial fermentation, part of the polysaccharide can be degraded , protein, fat and other macromolecular substances, generate small molecular substances such as organic acids and soluble polypeptides, accumulate beneficial metabolites, eliminate anti-nutritional factors, improve the digestibility of shrimp feed, and promote a series of functional strains in the shrimp intestine and aquaculture water. The population structure of beneficial and dominant microorganisms in the intestinal tract and water body of shrimps is stabilized, and a long-term stable and effective breeding model for efficient and healthy shrimp farming is constructed.

Invention content:

The purpose of this invention is to provide a kind of preparation method of fermented shrimp compound feed, compared with common shrimp compound feed or conventionally fermented shrimp compound feed, the fermented shrimp compound feed obtained by the preparation method proposed by the present invention is, on the one hand, in biological Peptide content, protein, L-lactic acid content and probiotic colony counts and other indicators have been significantly improved. These improvements can greatly promote the digestion, absorption and utilization of feed by prawns, improve the intestinal health of prawns, and improve immunity of prawns. Survival rate, reduce the use of antibiotics in shrimp farming, and reduce environmental pollution; on the other hand, feeding fermented shrimp compound feed can promote the proliferation of a series of functional strains in the shrimp intestine and aquaculture water, and stabilize the beneficial and dominant microbial populations in the shrimp intestine and water body. structure, build a long-term stable and effective breeding model for efficient and healthy shrimp farming; finally, fermentation of Aspergillus niger can produce multiple enzymes, enzymatically hydrolyze the anti-nutritional factors of the fermentation raw material soybean meal, and degrade into digestible nutrients, and the fermentation raw materials are pretreated and fermented. It is directly fed without granulation, and the production process is simple.

The fermented shrimp compound feed of the present invention is prepared by the following method:

(1) Raw materials of solid fermentation medium: based on the total mass fraction of 100%, including 10% to 30% of corn residue, 30% to 55% of soybean meal, 10% to 30% of fish meal, and 0.5% to 1.5% of shrimp compound premix feed , the remainder is bran, and the above-mentioned raw material components are mixed uniformly to obtain a mixture, and water is added until the weight water content of the mixture is 25% to 35% to obtain a solid fermentation medium;

(2) the aspergillus niger liquid, the bacillus subtilis bacterial liquid and the bacillus licheniformis bacterial liquid are inoculated into the solid fermentation medium and mixed evenly and then carry out aerobic fermentation to obtain the solid fermentation medium after the aerobic fermentation;

(3) adding sterile water to keep the water weight content of the solid fermentation medium after the aerobic fermentation at 25% to 35%, and inoculating the Lactobacillus reuteri bacterial liquid, Lactobacillus plantarum bacterial liquid and Saccharomyces cerevisiae liquid into The solid fermentation medium after the aerobic fermentation is evenly mixed and then anaerobic fermentation is carried out. After the fermentation is completed, the fermented shrimp compound feed is obtained.

Described aspergillus niger liquid, Bacillus subtilis bacterial liquid and Bacillus licheniformis bacterial liquid are inoculated into the solid fermentation medium and mixed evenly and then carry out aerobic fermentation, preferably the Aspergillus niger liquid, Bacillus subtilis bacterial liquid and lichen The Bacillus bacteria liquid is inoculated into the solid fermentation medium in a volume ratio of 6-8:1-2:1-2 and mixed evenly, and then aerobic fermentation is carried out until the material is covered with white mycelium, which is regarded as the end point of aerobic fermentation. Fermentation for 24 to 72 hours at a temperature of 28 to 32 degrees Celsius. It is further preferred that the inoculation amount is inoculated according to the total mass of the three kinds of bacteria: the mass of the solid fermentation medium=1:10.

The said Lactobacillus reuteri bacterial liquid, Lactobacillus plantarum bacterial liquid and Saccharomyces cerevisiae bacterial liquid are inoculated into the solid fermentation medium after aerobic fermentation and mixed evenly to carry out anaerobic fermentation, preferably Lactobacillus reuteri The mixed bacterial liquid of bacterial liquid, Lactobacillus plantarum bacterial liquid and Saccharomyces cerevisiae liquid is inoculated into the solid fermentation medium after aerobic fermentation at a volume ratio of 5 to 7:5 to 7:1, and then anaerobic fermentation is carried out after mixing evenly. . It is further preferred that the inoculation amount is inoculated according to the total mass of the three kinds of bacteria: the mass of the solid fermentation medium=1:10. The anaerobic fermentation conditions are as follows: the temperature is 28°C to 35°C, the time is 36 to 72 hours, and after the fermentation is completed, the fermented shrimp compound feed is obtained.

The 1000kg described prawn compound premix feed is prepared from the raw materials in the following mass ratios: vitamin A 1.0kg, vitamin D3 0.22kg, vitamin K3 2.0kg, dl-α-tocopheryl acetate 8.08kg, vitamin B11. 06kg, vitamin B2 1.52kg, vitamin B6 2.03kg, vitamin B12 0.20kg, niacinamide 7.12kg, D-calcium pantothenate 3.16kg, folic acid 0.73kg, D-biotin 0.92kg, L-ascorbic acid-2-phosphate 15.29kg , inositol 8.29kg, ethoxyquinoline 0.52kg, magnesium sulfate monohydrate 45.50kg, ferrous sulfate monohydrate 18.0kg, zinc sulfate monohydrate 12.49kg, copper sulfate pentahydrate 3.0kg, manganese sulfate monohydrate 2.72kg, 1% cobalt sulfate 1.0kg, 1% calcium iodate 1.5kg, 1% sodium selenite 1.0kg, and the rest are zeolite powder and rice husk powder in equal weight.

The lowest concentration of each bacterial solution is: Aspergillus niger solution 2.0×10 ⁷ cfu/mL, Bacillus subtilis solution 1.0×10 ⁹ cfu/mL, Bacillus licheniformis solution 8.0×10 ⁸ cfu/mL, Lactobacillus plantarum solution 0.8×10 ⁹ cfu/mL, Lactobacillus reuteri strain 1.0×10 ⁸ cfu/mL, Saccharomyces cerevisiae strain 7.0×10 ⁸ cfu/mL.

The fermented shrimp compound feed proposed by the present invention mainly uses corn residue, soybean meal, fish meal, bran, trace element additives, and vitamin additives as fermentation raw materials. Carry out aerobic fermentation to produce a large number of enzymes such as cellulase, xylanase, protease, amylase, saccharification enzyme, lipase, glucanase, etc., so as to decompose the starch, crude fiber and xylan of the fermentation raw materials into fermentable Sugar and protein are decomposed into biological peptides, etc., and then add Lactobacillus reuteri, Lactobacillus plantarum, Saccharomyces cerevisiae and other anaerobic fermentation to continue to decompose the macromolecular substances of fermentation raw materials, and produce L-lactic acid, reuterin and other inhibitory substances. Bacterial substances and aromatic odor, and lactic acid lowers pH, effectively inhibiting the growth of harmful bacteria.

The present invention utilizes Aspergillus niger, Bacillus subtilis, Bacillus licheniformis to produce abundant cellulase, xylanase, protease, amylase, saccharification enzyme, lipase, glucanase, etc., to comprehensively synergistically decompose the basic fermentation raw materials , macromolecular proteins are decomposed into peptides and amino acids, cellulose is decomposed into fermentable sugars, starch is decomposed into fermentable glucose, and xylan is decomposed into xylose, glucan, etc. After aerobic fermentation, the nutrients in the shrimp compound feed are released in large quantities, which can fully provide the necessary nutrients for other microorganisms such as lactic acid bacteria and yeast with weak decomposition ability. After the aerobic fermentation, the mixed strains consisting of Lactobacillus reuteri, Lactobacillus plantarum, and Saccharomyces cerevisiae were immediately inoculated according to a certain proportion, and then anaerobic fermentation was performed. After aerobic and anaerobic fermentation, the fermented shrimp compound feed was added. The crude protein content is 45% to 51%, the biological peptide content is 15% to 17%, the L-lactic acid content is 60 to 80 mg/g, the pH is 4.1 to 4.5, the total number of Bacillus > 1×10 ⁹ cfu/g, the total number of lactic acid bacteria ＞1×10 ¹⁰ cfu/g; in addition, fermentation metabolites contain a large amount of digestive enzymes, bacteriostatic substances and immune enhancing factors. After fermentation of probiotics, the anti-nutritional factors contained in plant protein are basically degraded. It has good sour aroma and wine aroma, and its pH is between 4.0 and 4.5, which is conducive to preservation.

The preparation method adopts the process of mixed fermentation of solid and liquid, preferably produces abundant enzymes and can co-grow Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, and carries out joint fermentation, and through aerobic and anaerobic two-step fermentation, to give full play to Aspergillus niger And Bacillus subtilis has the advantages of abundant enzymes, so that the basic fermentation raw materials are fully decomposed, macromolecular proteins are decomposed into peptides and amino acids, cellulose is decomposed into fermentable sugars, starch is decomposed into fermentable glucose, and xylan is decomposed It is xylose, glucan, etc., so that the growth and reproduction of Lactobacillus reuteri in the anaerobic stage can be supported by a large amount of nutrients. The biological peptide content, the L-lactic acid content, the number of colonies and other indicators can reflect that the growth and metabolism of microorganisms in the aerobic stage and the anaerobic stage in the preparation method proposed by the present invention are very vigorous.

Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, Lactobacillus reuteri, Lactobacillus plantarum, Saccharomyces cerevisiae of the present invention All of them are conventional strains in the field, and can be purchased from major collection centers or strain sales organizations.

Unless otherwise specified, the definitions of terms involved in the present invention have the same meanings as commonly understood by those skilled in the art.

Compared with the prior art, the present invention has the following advantages:

(1) The fermented shrimp compound feed proposed by the present invention has a high content of biological peptides, and the peptides can be directly absorbed by the animal intestines, and its digestion and absorption rate is higher than that of amino acids, and the absorption of peptides does not compete with the absorption of amino acids, and has biological The combination of active small peptides and trace elements can greatly promote the absorption and utilization of trace elements by aquatic animals, thereby promoting the growth of aquatic animals, reducing costs and environmental pollution. The digestion and absorption of prawns can reduce the burden on the gastrointestinal tract of prawns, enhance the disease resistance of prawns, and improve the utilization of trace mineral elements, which greatly improves the adverse effects of high-protein feed on the growth and development of prawns.

(2) The fermentation and metabolism of the fermented shrimp compound feed proposed by the present invention produces a large amount of digestive enzymes, L-lactic acid, bacteriostatic substances, etc., which can improve the nutritional level and utilization rate of the feed, inhibit the reproduction of harmful bacteria in the shrimp intestinal tract, and enhance the resistance of the shrimp. On the other hand, Lactobacillus reuteri and its metabolites have a certain repairing effect on the intestinal tract of shrimp; the use of fermented shrimp compound feed can improve the intestinal environment of shrimp, inhibit the growth of harmful bacteria, reduce the use of antibiotics and improve the feed of aquatic animals. Conversion rate, etc. have unique effects.

(3) The fermented shrimp compound feed proposed by the present invention has high content of Bacillus subtilis and Bacillus licheniformis, which can quickly degrade organic debris such as residual bait and feces in water, reduce ammonia nitrogen, hydrogen sulfide, etc. in the water environment, and avoid organic debris. Waste accumulates in the pool to maintain the balance of water ecology.

(4) The preparation method of the fermented shrimp compound feed proposed by the present invention adopts the technology of mixed fermentation of solid and liquid, and preferably produces abundant enzymes and can co-grow Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, and carries out joint fermentation, and through aerobic And anaerobic two-step fermentation, give full play to the advantages of rich enzymes produced by Aspergillus niger and Bacillus subtilis, so that the basic fermentation raw materials are fully decomposed, macromolecular proteins are decomposed into peptides and amino acids, and cellulose is degraded into fermentable sugars and starches It is decomposed into fermentable glucose, and xylan is decomposed into xylose, glucan, etc., so that the growth and reproduction of Lactobacillus reuteri in the anaerobic stage can be supported by a large amount of nutrients.

Detailed ways:

For the purpose of describing the invention, the following examples are set forth. It is to be understood, however, that the invention is not limited to these examples, but merely provides methods of practicing the invention.

Example 1:

(1) Preparation of liquid fermentation strains

a. Preparation of Aspergillus niger (purchased from the Guangdong Provincial Microbial Culture Collection Center, the preservation number is GIM 3.576) fermentation broth: place the slant strain of Aspergillus niger in an incubator, activate at 28°C for 24h, pick the The colony was inoculated into a 250mL conical flask containing 100mL PDA liquid medium for fermentation culture. The culture conditions were: temperature 28°C, time 24h, to obtain Aspergillus niger fermentation broth, the content of Aspergillus niger in Aspergillus niger fermentation broth was (2.0～4.0 )×10 ⁷ cfu/mL.

Among them, the formula of PDA liquid culture medium is: peeled potatoes 200g, chopped and boiled juice filtered to remove residue, add 20g of sucrose, 10g of peptone, 18g of agar, add water to make the volume to 1L, and adjust the pH to 7.0.

b. Preparation of Bacillus subtilis (purchased from the Guangdong Provincial Microorganism Culture Collection, the preservation number is GIM1.372) fermentation broth: pick the Bacillus subtilis and inoculate it into the LB medium, activate it on a shaking table at 37°C for 16 hours, and rotate the shaking table for 16 hours. Then, the activated bacterial liquid was inoculated into the primary seed medium according to the volume ratio of 3% inoculum for fermentation culture, and the culture conditions were: temperature 37 ° C, time 24 h, to obtain Bacillus subtilis fermentation broth, The content of Bacillus subtilis in the Bacillus subtilis fermentation broth was (1.0～1.8)×10 ⁹ cfu/mL.

Wherein, the formula of the LB medium (Luria-Bertani medium) is as follows: 10 g of tryptone, 5 g of yeast powder, and 10 g of sodium chloride are added to 1 L of distilled water to adjust the pH to 7.0.

The first-class seed medium is: add glucose 5g, corn starch 15g, soybean meal powder 20g, yeast powder 5g, magnesium sulfate 2g, dipotassium hydrogen phosphate 2g, manganese sulfate 0.3g, calcium carbonate 2.4g into distilled water, and then use distilled water Dilute to 1L and adjust pH to 7.0.

c. Preparation of Bacillus licheniformis (purchased from the Guangdong Provincial Microbial Culture Collection Center, the preservation number is GIM1.11) fermentation broth: pick Bacillus licheniformis and inoculate it into the LB medium, activate it on a shaking table at 37°C for 24 hours, and rotate the shaking table for 24 hours. Then, the activated bacterial liquid was inoculated into the primary seed medium according to the volume ratio of 3% inoculum for fermentation culture. The content of Bacillus licheniformis in the fermentation broth of Bacillus licheniformis was (8.0～8.8)×10 ⁸ cfu/mL.

Wherein, the formula of the LB medium (Luria-Bertani medium) is as follows: 10 g of tryptone, 5 g of yeast powder, and 10 g of sodium chloride are added to 1 L of distilled water to adjust the pH to 7.0.

The first-class seed medium is: add 5 g of glucose, 10 g of sucrose, 20 g of peptone, 3 g of yeast powder, 2 g of magnesium sulfate, 2.5 g of dipotassium hydrogen phosphate, and 5 g of ammonium chloride into distilled water, and then dilute to 1 L with distilled water. pH7.0.

d. Preparation of fermentation broth of Lactobacillus plantarum (purchased from the Guangdong Provincial Microbial Culture Collection Center, the preservation number is GIM1.191): inoculate the Lactobacillus plantarum into the MRS medium, stand at 37°C for activation for 24h, and then activate the The bacterial liquid of 5% by volume is inoculated into the first-class seed medium for fermentation culture, and the culture conditions are: stand still, temperature 37 ℃, time 18 ~ 20h, to obtain Lactobacillus plantarum fermentation broth, Lactobacillus plantarum The content of Lactobacillus plantarum in the fermentation broth was (0.8～1.5)×10 ⁹ cfu/mL.

The formula of MRS medium is: 10g peptone, 10g beef extract, 5g yeast extract, 2g diammonium hydrogen citrate, 20g glucose, 1mL Tween 80, 5g sodium acetate, 2g dipotassium hydrogen phosphate, 0.5g magnesium sulfate , 0.25 g of manganese sulfate was added to the water, and then the volume was adjusted to 1000 mL with distilled water, and the pH was adjusted to 6.2.

The first-class seed medium is: add 5g of glucose, 5g of molasses, 10g of peptone, 5g of yeast extract, 5g of sodium acetate, 2g of dipotassium hydrogen phosphate, and 0.5g of manganese sulfate into distilled water, then dilute to 1.0L with distilled water, adjust pH6.8.

e. Preparation of Lactobacillus reuteri (purchased from Guangdong Provincial Microbial Culture Collection Center, preservation number is CICC6118) fermentation broth: inoculate Lactobacillus reuteri into MRS medium, stand at 37°C for activation for 24h, and then The activated bacterial liquid is inoculated into the primary seed medium for fermentation culture according to the inoculation amount of 5% by volume. The culture conditions are: standing, temperature 37 ° C, time 20-22 h, to obtain Lactobacillus reuteri fermentation liquid , the content of Lactobacillus reuteri in the fermentation broth of Lactobacillus reuteri was (1.0～1.4)×10 ⁸ cfu/mL.

The formula of MRS medium is: 10g peptone, 10g beef extract, 5g yeast extract, 2g diammonium hydrogen citrate, 20g glucose, 1mL Tween 80, 5g sodium acetate, 2g dipotassium hydrogen phosphate, 0.5g magnesium sulfate , 0.25 g of manganese sulfate was added to distilled water, and then the volume was adjusted to 1000 mL with distilled water, and the pH was adjusted to 6.2.

The first-class seed medium is: add glucose 5g, molasses 5g, peptone 10g, yeast extract 5g, sodium acetate 5g, dipotassium hydrogen phosphate 2g, and manganese sulfate 0.5g into distilled water, then dilute to 1L with distilled water, and adjust pH to 6 .8.

f, preparation of Saccharomyces cerevisiae (purchased from Guangdong Provincial Microorganism Culture Collection Center, preservation number is GIM2.167) fermentation broth: pick Saccharomyces cerevisiae slant strain and inoculate it into PDA liquid medium, activate at 28° C. shaker for 24h, shake The bed rotation speed is 180r/min; then the activated bacterial liquid is inoculated into the first-class seed medium according to the volume ratio of 2% inoculum for fermentation culture. , the content of Saccharomyces cerevisiae in the fermentation broth was (7.0～8.0)×10 ⁸ cfu/mL.

Wherein, the formula of PDA liquid culture medium is the same as above.

The first-class seed medium is: add 20 g of glucose, 20 g of sucrose, 30 g of soy peptone, 10 g of yeast powder, 5 g of dipotassium hydrogen phosphate, and 5 g of urea into distilled water, and then dilute to 1.0 L with distilled water, pH 5.8 to 6.2.

(2) The raw materials of the solid fermentation medium: 10% corn residue, 55% soybean meal, 10% fish meal, 0.5% prawn compound premix feed, 24.5% bran, the above components are uniformly mixed according to the above component contents to obtain a mixture, and then Water was added to the mixture until its weight water content was 25% to obtain a solid fermentation medium.

Described prawn compound premixed feed (embodiments 2 and 3 are the same), every 1000kg is prepared from the raw materials of the following mass ratios: vitamin A 1.0kg, vitamin D3 0.22kg, vitamin K3 2.0kg, dl-α -Tocopheryl acetate 8.08kg, vitamin B1 1.06kg, vitamin B2 1.52kg, vitamin B6 2.03kg, vitamin B12 0.20kg, niacinamide 7.12kg, D-calcium pantothenate 3.16kg, folic acid 0.73kg, D-biotin 0.92 kg, L-ascorbic acid-2-phosphate 15.29kg, inositol 8.29kg, ethoxyquinoline 0.52kg, magnesium sulfate monohydrate 45.50kg, ferrous sulfate monohydrate 18.0kg, zinc sulfate monohydrate 12.49kg, pentahydrate 3.0kg of copper sulfate, 2.72kg of manganese sulfate monohydrate, 1.0kg of 1% cobalt sulfate, 1.5kg of 1% calcium iodate, 1.0kg of 1% sodium selenite, and the rest are zeolite powder and rice husk powder in equal parts. The raw materials are evenly mixed to obtain the composite premixed feed for prawns. Wherein, the 1% cobalt sulfate, 1% calcium iodate and 1% sodium selenite were all purchased from Guangzhou Zhizhuan Feed Co., Ltd.

(3) aerobic fermentation stage: the liquid fermentation strains prepared in step (1) are inoculated to a ratio of 6:1:1 according to the volume ratio of Aspergillus niger fermentation broth, Bacillus subtilis fermentation broth and Bacillus licheniformis fermentation broth In the solid fermentation medium that step (2) obtains, the amount of inoculation is that the mass ratio is 10% (i.e. Aspergillus niger fermentation broth, Bacillus subtilis fermentation broth and Bacillus licheniformis three kinds of bacteria total mass every 1g, inoculated into the solid fermentation culture of 10g. base), placed in an open container for aerobic fermentation, the fermentation conditions are: temperature 28 ℃, time 24h, to obtain a solid fermentation medium after aerobic fermentation, the material covered with white mycelium is regarded as the end point of aerobic fermentation, The next stage of fermentation is then carried out.

(4) anaerobic fermentation stage: adding sterile water to keep the water content of the solid fermentation medium after the aerobic fermentation at 25% of the weight water content, the prepared liquid fermentation strains are according to the Lactobacillus reuteri fermentation broth, and the plant The volume ratio of Lactobacillus fermented liquid and Saccharomyces cerevisiae fermented liquid is the ratio of 5:5:1, inoculated into the solid fermentation medium after aerobic fermentation, and the inoculation amount is 10% (i.e. Lactobacillus reuteri fermentation liquid, plant Lactobacillus fermented liquid and Saccharomyces cerevisiae fermented liquid each 1g of the total mass of three bacteria, inoculated into 10g of solid fermentation medium after aerobic fermentation), placed in a sealed airtight container for anaerobic fermentation, and the fermentation conditions are: temperature 28 ℃, time 36h, after the fermentation is completed, the fermented shrimp compound feed is obtained.

Measure the crude protein content, biopeptide content, L-lactic acid content, total number of bacillus, total number of lactic acid bacteria and other indicators of the unfermented shrimp compound feed (that is, the solid fermentation medium of step (2)) and the fermented shrimp compound feed, as shown in Table 1 shown.

Table 1

project Unfermented shrimp compound feed Example 1 Preparation of fermented shrimp compound feed Crude protein (%) 36 45 Biopeptides (%) 0.4 15 L-lactic acid (mg/g) 0.5 60 Total Bacillus 1.2×10³ 1.0×10⁹ The total number of lactic acid bacteria 0 1.4×10¹⁰

Example 2:

(1) Prepare liquid fermentation strains (Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, Lactobacillus reuteri, Lactobacillus plantarum, Saccharomyces cerevisiae) according to the method of Example 1.

(2) Weigh the raw materials according to the following mass fractions: 20% of corn residue, 45% of soybean meal, 20% of fish meal, 0.8% of prawn compound premix feed, and 14.2% of bran, and mix the above components uniformly to obtain a mixture. Water was added until its weight water content was 30% to obtain a solid fermentation medium.

(3) aerobic fermentation stage: the liquid fermentation strain prepared in step (1) is inoculated into the Aspergillus niger fermentation broth, the Bacillus subtilis fermentation broth and the Bacillus licheniformis fermentation broth in a volume ratio of 7:2:1 The solid fermentation medium prepared by step (2), the inoculum amount is 10% by mass (the total mass of three kinds of bacteria: the quality of the solid fermentation medium=1:10), placed in an open container and carried out aerobic fermentation, fermentation conditions As follows: the temperature is 30°C and the time is 48h to obtain the solid fermentation medium after aerobic fermentation. The material covered with white mycelium is regarded as the end point of aerobic fermentation, and then the next stage of fermentation is carried out.

(4) anaerobic fermentation stage: adding sterile water to keep the water content of the solid fermentation medium after the aerobic fermentation at 30% of the water content by weight, the prepared liquid fermentation strains were prepared according to the Lactobacillus reuteri fermentation broth, and the plants The volume ratio of Lactobacillus fermented liquid and Saccharomyces cerevisiae fermented liquid is that the ratio of 6:6:1 is inoculated into the solid fermentation medium after the aerobic fermentation of step (3), and the inoculation amount is 10% (the total mass of three kinds of bacteria: The quality of the solid fermentation medium after aerobic fermentation = 1:10), placed in a sealed airtight container for anaerobic fermentation, the fermentation conditions are: temperature 32 ° C, time 54h, after the fermentation is completed, the fermented shrimp is obtained. feed.

Measure indicators such as feed crude protein content, biopeptide content, L-lactic acid content, total number of Bacillus, total number of lactic acid bacteria, etc. shown.

Table 2

project Unfermented shrimp compound feed Embodiment 2 makes fermented shrimp compound feed Crude protein (%) 37.6 48.5 Biopeptides (%) 0.6 16 L-lactic acid (mg/g) 0.62 67 Total Bacillus 1.1×10³ 2.4×10⁹ The total number of lactic acid bacteria 0 1.8×10¹⁰

Example 3:

(1) Prepare liquid fermentation strains (Aspergillus niger, Bacillus subtilis, Bacillus licheniformis, Lactobacillus reuteri, Lactobacillus plantarum, Saccharomyces cerevisiae) according to the method of Example 1.

(2) Weigh the raw materials according to the following mass fractions: 30% of corn residue, 30% of soybean meal, 30% of fish meal, 1.5% of shrimp compound premix feed, and 8.5% of bran. Water was added to the mixture until its water content by weight was 35% to obtain a solid fermentation medium.

(3) Aerobic fermentation stage: the liquid fermentation strain prepared in step (1) is inoculated into the step according to the volume ratio of Aspergillus niger fermentation broth, Bacillus subtilis fermentation broth and Bacillus licheniformis fermentation broth in a ratio of 8:1:2 (2) the prepared solid fermentation medium, the inoculum amount is 10% by mass (the total mass of three kinds of bacteria: the quality of the solid fermentation medium=1:10), is placed in an open container to carry out aerobic fermentation, and the fermentation conditions are : The temperature is 32°C and the time is 72h to obtain the solid fermentation medium after aerobic fermentation. The material covered with white mycelium is regarded as the end point of aerobic fermentation, and then the next stage of fermentation is carried out.

(4) anaerobic fermentation stage: adding sterile water to keep the water content of the solid fermentation medium after the aerobic fermentation at 35% of the water content by weight, the prepared liquid fermentation strains were prepared according to the Lactobacillus reuteri fermentation broth, and the plant The volume ratio of Lactobacillus fermented liquid and Saccharomyces cerevisiae fermented liquid is that the ratio of 7:7:1 is inoculated into the solid fermentation medium after the aerobic fermentation of step (3), and the inoculation amount is 10% (the total mass of three kinds of bacteria: The quality of the solid fermentation medium after aerobic fermentation = 1:10), placed in a sealed airtight container for anaerobic fermentation, the fermentation conditions are: temperature 35 ° C, time 72 h, after the fermentation is completed, the fermented shrimp is obtained. feed.

Measure the crude protein content, biopeptide content, L-lactic acid content, total number of bacillus, total number of lactic acid bacteria and other indicators of unfermented shrimp compound feed (that is, the solid fermentation medium of step (2)) and fermented shrimp compound feed, as shown in Table 3 shown.

table 3

project Unfermented shrimp compound feed The fermented shrimp compound feed prepared in Example 3 Crude protein (%) 38 51 Biopeptides (%) 0.8 17 L-lactic acid (mg/g) 1.0 80 Total Bacillus 1.1×10³ 1.5×10⁹ The total number of lactic acid bacteria 0 2.0×10¹⁰

It can be drawn from Tables 1 to 3 that the fermented shrimp compound feed prepared in Examples 1 to 3 is compared with the unfermented shrimp compound feed (ie, the solid fermentation medium of each step (2)), and the crude protein content is determined by 36%～38% to 45%～51%, biopeptide content from 0.4%～0.8% to 15%～17%, L-lactic acid content from 0.5～1mg/g to 60～80mg/g, pH The value decreased from 6.3 to 6.5 to 4.1 to 4.5, the total number of Bacillus > 1 × 10 ⁹ cfu/g, the total number of lactic acid bacteria > 1 × 1010 cfu/g.

Claims (2)

1. A preparation method of a fermented prawn compound feed is characterized by comprising the following steps:

(1) preparing a solid fermentation culture medium raw material: the raw material components comprise 10-30% of corn grit, 30-55% of soybean meal, 10-30% of fish meal, 0.5-1.5% of prawn composite premixed feed and the balance of bran, the raw material components are uniformly mixed to obtain a mixture, water is added until the water content of the mixture is 25-35%, and a solid fermentation culture medium with the pH value of 6.3-6.5 is obtained;

(2) inoculating Aspergillus niger liquid, Bacillus subtilis liquid and Bacillus licheniformis liquid into a solid fermentation culture medium in a volume ratio of 6-8: 1-2, uniformly mixing, and performing aerobic fermentation, wherein the mass ratio of the total mass of the Aspergillus niger liquid, the Bacillus subtilis liquid and the Bacillus licheniformis liquid to the solid fermentation culture medium is 1:10, and the aerobic fermentation conditions are as follows: fermenting for 24-72 h at the temperature of 28-32 ℃;

(3) adding sterile water to keep the water content of the solid fermentation culture medium after aerobic fermentation to be 25-35%, inoculating the mixed bacterial liquid of the lactobacillus reuteri bacterial liquid, the lactobacillus plantarum bacterial liquid and the saccharomyces cerevisiae bacterial liquid into the solid fermentation culture medium after the aerobic fermentation in a volume ratio of 5-7: 1, uniformly mixing, and then carrying out anaerobic fermentation, thus obtaining the fermented prawn compound feed with the pH value of 4.1-4.5 after the fermentation is finished, wherein the mass ratio of the total mass of the lactobacillus reuteri bacterial liquid, the lactobacillus plantarum bacterial liquid and the saccharomyces cerevisiae bacterial liquid to the solid fermentation culture medium is 1:10, and the anaerobic fermentation conditions are as follows: the temperature is 28-3 ℃, and the time is 36-72 h;

the concentration of each bacterial liquid of the aspergillus niger bacterial liquid, the bacillus subtilis bacterial liquid, the bacillus licheniformis bacterial liquid, the lactobacillus reuteri bacterial liquid, the lactobacillus plantarum bacterial liquid and the saccharomyces cerevisiae bacterial liquid is the lowest: aspergillus niger liquid 2.0X 10⁷cfu/mL, 1.0X 10 Bacillus subtilis liquid⁹cfu/mL, Bacillus licheniformis liquid 8.0 × 10⁸cfu/mL, Lactobacillus plantarum bacterial liquid 0.8 × 10⁹cfu/mL, Lactobacillus reuteri bacterial liquid 1.0 × 10⁸cfu/mL, Saccharomyces cerevisiae bacterial liquid 7.0 × 10⁸cfu/mL; the compound premixed feed for prawns comprises, per 1000 kg: 1.0kg of vitamin A, 30.22 kg of vitamin D, 32.0 kg of vitamin K, 8.08kg of dl-a-tocopheryl acetate, 11.06kg of vitamin B, 21.52kg of vitamin B, 62.03 kg of vitamin B, 120.20 kg of vitamin B, 7.12kg of nicotinamide, 3.16kg of D-calcium pantothenate, 0.73kg of folic acid, 0.92kg of D-biotin, 15.29kg of L-ascorbic acid-2-phosphate, 8.29kg of inositol, 0.52kg of ethoxyquinoline, 0.52kg of magnesium sulfate monohydrate45.50kg, 18.0kg of ferrous sulfate monohydrate, 12.49kg of zinc sulfate monohydrate, 3.0kg of copper sulfate pentahydrate, 2.72kg of manganese sulfate monohydrate, 1.0kg of 1% cobalt sulfate, 1% of calcium iodate, 1.5kg of 1% sodium selenite, and the balance of zeolite powder and half of rice hull powder in mass.

2. A fermented prawn compound feed prepared according to the preparation method of claim 1.