CN114304379A - Preparation method of fermented feed containing compound microbial agent - Google Patents
Preparation method of fermented feed containing compound microbial agent Download PDFInfo
- Publication number
- CN114304379A CN114304379A CN202210024704.7A CN202210024704A CN114304379A CN 114304379 A CN114304379 A CN 114304379A CN 202210024704 A CN202210024704 A CN 202210024704A CN 114304379 A CN114304379 A CN 114304379A
- Authority
- CN
- China
- Prior art keywords
- fermented feed
- feed containing
- microbial agent
- compound microbial
- fermentation
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 230000000813 microbial effect Effects 0.000 title claims abstract description 35
- 150000001875 compounds Chemical class 0.000 title claims abstract description 32
- 238000002360 preparation method Methods 0.000 title claims abstract description 14
- 241000194031 Enterococcus faecium Species 0.000 claims abstract description 37
- 244000063299 Bacillus subtilis Species 0.000 claims abstract description 33
- 235000014469 Bacillus subtilis Nutrition 0.000 claims abstract description 33
- 240000001046 Lactobacillus acidophilus Species 0.000 claims abstract description 33
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims abstract description 33
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims abstract description 33
- 244000046052 Phaseolus vulgaris Species 0.000 claims abstract description 30
- 235000010627 Phaseolus vulgaris Nutrition 0.000 claims abstract description 30
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 19
- 244000017020 Ipomoea batatas Species 0.000 claims abstract description 17
- 235000002678 Ipomoea batatas Nutrition 0.000 claims abstract description 17
- 240000003183 Manihot esculenta Species 0.000 claims abstract description 17
- 235000016735 Manihot esculenta subsp esculenta Nutrition 0.000 claims abstract description 17
- 244000061456 Solanum tuberosum Species 0.000 claims abstract description 17
- 235000002595 Solanum tuberosum Nutrition 0.000 claims abstract description 17
- 238000002156 mixing Methods 0.000 claims abstract description 7
- 239000002131 composite material Substances 0.000 claims abstract 4
- 238000000855 fermentation Methods 0.000 claims description 45
- 230000004151 fermentation Effects 0.000 claims description 45
- 239000001963 growth medium Substances 0.000 claims description 41
- 239000007787 solid Substances 0.000 claims description 35
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 25
- 239000007788 liquid Substances 0.000 claims description 23
- 230000001580 bacterial effect Effects 0.000 claims description 7
- 239000002699 waste material Substances 0.000 claims description 5
- 238000010564 aerobic fermentation Methods 0.000 claims description 4
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 2
- 238000013329 compounding Methods 0.000 claims description 2
- 239000001301 oxygen Substances 0.000 claims description 2
- 229910052760 oxygen Inorganic materials 0.000 claims description 2
- 239000002054 inoculum Substances 0.000 claims 5
- 239000002075 main ingredient Substances 0.000 claims 1
- 239000002609 medium Substances 0.000 claims 1
- 150000001413 amino acids Chemical class 0.000 abstract description 12
- 235000019750 Crude protein Nutrition 0.000 abstract description 11
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 239000002994 raw material Substances 0.000 abstract description 7
- 230000008901 benefit Effects 0.000 abstract description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000010563 solid-state fermentation Methods 0.000 abstract description 3
- 235000019764 Soybean Meal Nutrition 0.000 abstract 1
- 239000002068 microbial inoculum Substances 0.000 abstract 1
- 230000000050 nutritive effect Effects 0.000 abstract 1
- 235000012015 potatoes Nutrition 0.000 abstract 1
- 239000004455 soybean meal Substances 0.000 abstract 1
- 238000009630 liquid culture Methods 0.000 description 18
- 241001465754 Metazoa Species 0.000 description 14
- 241000235342 Saccharomycetes Species 0.000 description 10
- 229920001817 Agar Polymers 0.000 description 6
- 239000008272 agar Substances 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 239000000843 powder Substances 0.000 description 6
- 241000894006 Bacteria Species 0.000 description 5
- 238000011081 inoculation Methods 0.000 description 5
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 4
- 239000001888 Peptone Substances 0.000 description 4
- 108010080698 Peptones Proteins 0.000 description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 235000015278 beef Nutrition 0.000 description 4
- 229940041514 candida albicans extract Drugs 0.000 description 4
- 239000000284 extract Substances 0.000 description 4
- 239000008103 glucose Substances 0.000 description 4
- 244000144972 livestock Species 0.000 description 4
- 235000019319 peptone Nutrition 0.000 description 4
- 239000006041 probiotic Substances 0.000 description 4
- 235000018291 probiotics Nutrition 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 239000000126 substance Substances 0.000 description 4
- 239000012138 yeast extract Substances 0.000 description 4
- 239000003674 animal food additive Substances 0.000 description 3
- 238000000576 coating method Methods 0.000 description 3
- 210000001035 gastrointestinal tract Anatomy 0.000 description 3
- 230000036039 immunity Effects 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 238000007789 sealing Methods 0.000 description 3
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 2
- 235000019728 animal nutrition Nutrition 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 244000052616 bacterial pathogen Species 0.000 description 2
- KLOIYEQEVSIOOO-UHFFFAOYSA-N carbocromen Chemical compound CC1=C(CCN(CC)CC)C(=O)OC2=CC(OCC(=O)OCC)=CC=C21 KLOIYEQEVSIOOO-UHFFFAOYSA-N 0.000 description 2
- 150000001720 carbohydrates Chemical class 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000011248 coating agent Substances 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 230000002708 enhancing effect Effects 0.000 description 2
- 239000003797 essential amino acid Substances 0.000 description 2
- 235000020776 essential amino acid Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000011187 glycerol Nutrition 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 2
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 2
- OQTQHQORDRKHFW-UHFFFAOYSA-L manganese(2+);sulfate;heptahydrate Chemical compound O.O.O.O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O OQTQHQORDRKHFW-UHFFFAOYSA-L 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 244000144977 poultry Species 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000001632 sodium acetate Substances 0.000 description 2
- 235000017281 sodium acetate Nutrition 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- 239000012137 tryptone Substances 0.000 description 2
- 239000004382 Amylase Substances 0.000 description 1
- 102000013142 Amylases Human genes 0.000 description 1
- 108010065511 Amylases Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 102000014150 Interferons Human genes 0.000 description 1
- 108010050904 Interferons Proteins 0.000 description 1
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 description 1
- 102000004882 Lipase Human genes 0.000 description 1
- 108090001060 Lipase Proteins 0.000 description 1
- 239000004367 Lipase Substances 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 239000004480 active ingredient Substances 0.000 description 1
- 239000000853 adhesive Substances 0.000 description 1
- 230000001070 adhesive effect Effects 0.000 description 1
- 235000019418 amylase Nutrition 0.000 description 1
- 238000003975 animal breeding Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 210000004211 gastric acid Anatomy 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 229940079322 interferon Drugs 0.000 description 1
- 230000031891 intestinal absorption Effects 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 235000019421 lipase Nutrition 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229930182817 methionine Natural products 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000003672 processing method Methods 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 235000019419 proteases Nutrition 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 235000015099 wheat brans Nutrition 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/80—Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
- Y02P60/87—Re-use of by-products of food processing for fodder production
Landscapes
- Fodder In General (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a preparation method of a fermented feed containing a compound microbial agent. The method comprises the following steps of adopting a bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and microzyme composite microbial inoculum to produce and prepare a composite microbial agent by uniformly mixing and fermenting sweet potato residues, cassava residues, potatoes, bean pulp, bran and other crude raw materials, and preparing a fermented feed containing the composite microbial agent, wherein the sweet potato residues: cassava residues: bran: potato residue: bean dregs: when the proportion of the soybean meal is 2:1:1:2:3:1, the total colony number in the fermented feed reaches 1010More than cfu/g, 24.39 +/-0.03 percent of crude protein, 4.34 +/-0.02 percent of amino acid and 23.44 +/-0.03 percent of water, and the best effect is achieved when the solid state fermentation time is 3-5 days, the temperature is 25-45 ℃, and the initial pH is 6.5. Compared with other prepared feeds, the method has the advantages of low cost, convenient production and operation, convenient use and raw material generationShort production period and the like, thereby improving the nutritive value and quality of the feed, obviously reducing the production cost and having obvious market application value.
Description
Technical Field
The invention belongs to the technical field of preparation of fermented feed of a compound microbial agent for feed, and particularly relates to a preparation method of fermented feed containing the compound microbial agent.
Background
The compound microbial agent for the feed is used as a novel feed additive, contains a large amount of probiotics, can maintain the balance of intestinal microorganisms, replace antibiotics, enhance the immunity, promote the health and the growth of animals, improve the feeding efficiency, reduce the feeding cost, reduce the antibiotic residue of animal products, inhibit drug-resistant pathogenic bacteria, improve the feed conversion rate, enhance the immune system of the animals and reduce the occurrence of diseases. In recent years, the compound microbial agent for feed is popularized, so that the feed conversion rate can be increased, the meat quality can be improved, considerable economic benefits are brought to the breeding industry, and the development of the animal husbandry is promoted.
The bacillus subtilis belongs to one of bacillus, is a mesophilic, aerobic and spore-producing gram-positive rod-shaped bacterium, is nontoxic and harmless to people and livestock, can secrete active ingredients such as amylase, protease, lipase and the like and produce antibacterial substances and the like, has broad-spectrum antibacterial activity, can produce a biological oxygen-depriving effect in animal intestinal tracts, provides an anaerobic environment for the growth of lactic acid bacteria, and plays an important role in the utilization, growth and disease prevention of animal nutrient substances, and meanwhile, the strain is listed in a feed additive catalogue by the agricultural rural part of China and is mainly applied to the aspect of microecologics; the lactobacillus acidophilus can be planted in the intestinal tracts of animals in a large amount, help food digestion and metabolism and promote the absorption and utilization of nutrient substances; meanwhile, the feed additive can compete with pathogenic bacteria, inhibit the reproduction of harmful bacteria, improve the environment in intestinal tracts, adjust the balance of gastrointestinal flora and improve the health level of animals; enterococcus faecium has the characteristics of strong heat resistance, strong drug resistance, strong adhesive force, gastric acid resistance and the like, and is widely researched and applied as a good feeding probiotic all the time due to the unique advantages of the enterococcus faecium in production and processing. A large number of researches prove that the enterococcus faecium fed by the feed can improve the daily gain of animals, enhance the immunity of organisms, reduce the diarrhea rate and the death rate and maintain the balance of intestinal microflora. At present, the enterococcus faecium microecological preparation is widely applied to the animal husbandry production as a substitute of antibiotics, and has good feeding prospect; the yeast fermentation culture has the advantages of obviously enhancing the titer of interferon, thereby enhancing the immune function of organisms, supplementing trace elements, secreting a large amount of protein and the like, and can improve the daily gain, improve the material-to-weight ratio, enhance the disease resistance of the organisms and improve the economic benefit in animal breeding. Therefore, in recent years, research on fermented feed containing the compound microbial agent and application thereof is more and more intensive, components such as protein, starch, cellulose and the like can be decomposed into easily digestible components such as small molecular protein, small peptide, free amino acid and the like, and the fermented feed is more and more beneficial to intestinal absorption of livestock and poultry. The feed rich in probiotics is used for a long time, so that the intestinal environment of livestock and poultry can be improved, the immunity of animals is enhanced, the obvious effects of promoting the growth of the animals are achieved, and the edible safety of livestock products is effectively guaranteed.
The prior report does not show a precedent that a plurality of crude raw materials are mixed together and fermented by probiotics to produce the feed of the compound microbial agent. The method combines the utilization of the crude raw materials with the microbial technology, can improve the treatment efficiency, improve the treatment effect, reduce the cost, further meet the requirement of resource utilization, and has wide market prospect.
Disclosure of Invention
The invention aims to provide a preparation method of a fermented feed containing a compound microbial agent, wherein the strains are bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycete compound bacteria, the compound bacteria are adopted to ferment various crude raw materials, and the fermented product is used as the feed.
In order to realize the purpose of the invention, the technical scheme is as follows:
a preparation method of a fermented feed containing a compound microbial agent comprises the steps of preparing four different types of bacterial strains into the compound microbial agent, fermenting and preparing the compound microbial agent by utilizing crude raw materials such as sweet potato residues, cassava residues, potato residues, bean dregs, bean pulp and bran, and producing and preparing the fermented feed containing the compound microbial agent in an optimized fermentation culture mode.
The method comprises the following steps:
s1, respectively carrying out liquid fermentation culture on bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast, and then compounding the bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the yeast into a compound microbial agent according to a proportion;
s2, inoculating the compound microbial agent of S1 into a solid culture medium for mixed fermentation; and (3) fermenting at the temperature of 25-45 ℃ for 3-5 days to obtain the fermented feed.
The bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the microzyme are prepared according to the following formula (1-6): (1-6): (1-6): (1-6) in a mass ratio.
In the fermentation, aerobic fermentation is carried out for 12 to 20 hours, and then the oxygen concentration is reduced by one time to continue the fermentation for 52 to 100 hours.
The main component in the solid culture medium is at least one of the processed sweet potato residue, the manioc waste, the bran, the potato residue, the bean dregs and the bean pulp.
Further preferably, the bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast are mixed according to a ratio of 1:1:1:1, in a mass ratio of 1.
The solid culture medium is prepared from the processed sweet potato residues, the processed cassava residues, the processed wheat bran, the processed potato residues, the processed bean dregs and the processed bean pulp in a mass ratio of (1-10): (1-10): (1-10): (1-10): (1-10): (1-10) mixing.
Respectively carrying out liquid fermentation culture on bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes, and inoculating the bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes into a solid fermentation culture medium according to the proportion of 2-15% (W/W) for fermentation.
The total bacterial colony number in the obtained fermented feed reaches 1010cfu/g or more.
Further preferably, the solid culture medium is prepared by mixing the processed sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp according to the mass ratio of 2:1:1:2:3: 1.
The fermented feed containing the compound microbial agent prepared by the preparation method.
In the fermented feed obtained in one embodiment of the invention, the total colony count reaches 1010cfu/g above, crude protein content 24.39 + -0.03%, amino acid 4.34 + -0.02%, and water content 23.44 + -0.03%.
The four strains adopted by the invention are all strains commonly used in the field, and can be directly obtained from commercial sources or obtained from a depository.
The method for fermenting the four strains comprises the following steps,
the method for separately fermenting the bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the microzyme in liquid state comprises the following steps,
(1) solid and liquid culture medium for strain
Liquid culture medium of bacillus subtilis: 10g of peptone, 5g of yeast powder and 10g of sodium chloride, and metering the volume to 1L; the solid culture medium is liquid culture medium added with 20% agar.
Liquid culture medium of yeast: tryptone 20g, yeast powder 10g and glucose 20g, wherein the volume is constant to 1L, and the pH value is 7.2-7.4; the solid culture medium is liquid culture medium added with 20% agar.
Liquid culture medium of lactobacillus acidophilus and enterococcus faecium: : 10.0 g of peptone, 10.0 g of beef extract (beef extract), 5.0 g of yeast extract (yeast extract or yeast powder), 20.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diammonium hydrogen citrate, 801.0 ml of tween-801.0 ml, 2.0 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate and 0.05 g of manganese sulfate heptahydrate, and the volume is fixed to 1000ml, and the pH value is 6.2-6.4. The culture time is 18-20 hours; the solid culture medium is liquid culture medium added with 20% agar.
The sterilization method comprises the following steps: sterilizing with high pressure steam at 115 deg.C for 15min for saccharides, and 121 deg.C for 20min for the rest.
(2) Solid plate culture of strain
Taking glycerin pipes of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes out of an ultra-low temperature refrigerator, transferring 100-300 microliters of bacterial liquid in an ultra-clean bench, and coating and culturing on a solid culture medium plate.
(3) Seed liquid culture
Respectively inoculating the four different strains which are cultured into respective seed liquid culture media, and putting the strains into a shaking table at the temperature of 30-37 ℃ for culturing, wherein the bacillus subtilis seed liquid culture is carried out at 200rpm for about 12 hours; culturing lactobacillus acidophilus and enterococcus faecium at 150rpm for about 18-20 h; the seed liquid of the yeast is cultured for about 24 hours at 200 rpm.
The method comprises the following steps of performing independent liquid fermentation on bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes, transferring the bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes into a solid culture medium, and performing fermentation, wherein seed liquids of four different strains which are independently subjected to liquid fermentation are prepared according to the volume ratio of (1-6): (1-6): (1-6): (1-6), inoculating the liquid strain after mixing and proportioning into a solid fermentation culture medium according to the proportion of 2-12% (V/W), subpackaging into plastic bags, opening the bag mouth at the temperature of 25-45 ℃, carrying out aerobic fermentation for 12-20 h, semi-sealing the bag mouth, and fermenting for 52-100 h to obtain the fermented feed containing the compound microbial agent.
In the solid fermentation preparation of the fermented feed containing the compound microbial agent, the total bacterial colony number reaches 1010cfu/g above, crude protein content 24.39 + -0.03%, amino acid 4.34 + -0.02%, and water content 23.44 + -0.03%. In the animal feed prepared by the solid fermentation, the content of crude protein and the content of amino acid are both higher than the essential amino acid required by the animal nutrition in the national standard (GB/T5916-.
Has the advantages that:
the product of the invention can change the production waste into valuable, realize the reutilization of waste resources, reduce the pollution of waste materials to the environment, greatly reduce the production cost, is economic and feasible, and has wide market application value, and the content of crude protein and the content of amino acid in the animal feed prepared by solid fermentation are both higher than the essential amino acid required by animal nutrition in the national standard (GB/T5916-2008).
Detailed Description
The invention will be better understood from the following examples. However, those skilled in the art will readily appreciate that the specific process conditions and results described in the examples are merely illustrative of the invention and should not, nor should they, limit the invention as described in detail in the claims.
The strains referred to in the following examples are: bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast;
bacillus subtilis CICC 24713 and Lactobacillus acidophilus CICC 20709 adopted in the embodiment are purchased from China center for culture Collection of industrial microorganisms;
the yeast CGMCC No.1189 is from the common microorganism center of China Committee for culture Collection of microorganisms;
enterococcus faecium is enterococcus faecium AS1.130, available from Jiangsu Biotech (Jiangsu) Inc.;
the sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp described in the following examples are purchased by oneself.
The sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp in the following examples have the following processing methods: after being uniformly mixed, the raw materials are physically crushed to ensure that the particles are uniform.
The method for separately fermenting the bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the microzyme in liquid state comprises the following steps,
liquid culture medium of bacillus subtilis: 10g of peptone, 5g of yeast powder and 10g of sodium chloride, and metering the volume to 1L; the solid culture medium is liquid culture medium added with 20% agar.
Liquid culture medium of yeast: tryptone 20g, yeast powder 10g and glucose 20g, wherein the volume is constant to 1L, and the pH value is 7.2-7.4; the solid culture medium is liquid culture medium added with 20% agar.
Liquid culture medium of lactobacillus acidophilus and enterococcus faecium: : 10.0 g of peptone, 10.0 g of beef extract (beef extract), 5.0 g of yeast extract (yeast extract or yeast powder), 20.0 g of glucose, 5.0 g of sodium acetate, 2.0 g of diammonium hydrogen citrate, 801.0 ml of tween-801.0 ml, 2.0 g of dipotassium hydrogen phosphate, 0.2 g of magnesium sulfate heptahydrate and 0.05 g of manganese sulfate heptahydrate, and the volume is fixed to 1000ml, and the pH value is 6.2-6.4. The culture time is 18-20 hours; the solid culture medium is liquid culture medium added with 20% agar.
The sterilization method comprises the following steps: sterilizing with high pressure steam at 115 deg.C for 15min for saccharides, and 121 deg.C for 20min for the rest.
Taking glycerin pipes of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes out of an ultra-low temperature refrigerator, transferring 100-300 microliters of bacterial liquid in an ultra-clean bench, and coating and culturing on a solid culture medium plate.
Respectively inoculating the four different strains which are cultured into respective seed liquid culture media, and putting the strains into a shaking table at the temperature of 30-37 ℃ for culturing, wherein the bacillus subtilis seed liquid culture is carried out at 200rpm for about 12 hours; culturing lactobacillus acidophilus and enterococcus faecium at 150rpm for about 18-20 h; the seed liquid of the yeast is cultured for about 24 hours at 200 rpm.
The method comprises the following steps of performing independent liquid fermentation on bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes, transferring the bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes into a solid culture medium, and performing fermentation, wherein seed liquids of four different strains which are independently subjected to liquid fermentation are prepared according to the volume ratio of (1-6): (1-6): (1-6): (1-6), inoculating the liquid strain after mixing and proportioning into a solid fermentation culture medium according to the proportion of 2-12% (V/W), subpackaging into plastic bags, opening the bag mouth at the temperature of 25-45 ℃, carrying out aerobic fermentation for 12-20 h, semi-sealing the bag mouth, and fermenting for 52-100 h to obtain the fermented feed containing the compound microbial agent.
Example 1
In the embodiment, the inoculation ratios of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast are considered, seed liquids of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast in different ratios are respectively inoculated into a solid fermentation culture medium with the ratio of sweet potato residue, cassava residue, bran, potato residue, bean dregs and bean pulp of 1:1:1:2:3:1 according to the inoculation amount of 4% (V/W), a fermentation experiment is carried out, the early stage is 20 hours, and the later stage is 52 hours. B, bacillus subtilis: lactobacillus acidophilus: enterococcus faecium: the yeast ratios were 1:0:0:1, 0:1:1:1, 1:1:1:0, 1:1:0:1, 1:0:1:1, 1:1:1:1, 2:2:2:2, respectively, and the total colony count, crude protein (dry basis), moisture (dry basis), and amino acid (dry basis) of the fermentation product were measured, and the results are shown in table 1.
TABLE 1 influence of different strain ratios on various physicochemical indices of fermentation products
Example 2
The invention inspects the proportion of the sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp in the solid fermentation culture medium, and the ratio of the bacillus subtilis: lactobacillus acidophilus: enterococcus faecium: adding liquid seed liquid with yeast ratio of 1:1:1:1 into solid fermentation culture medium with different ratio according to inoculation amount of 4% (V/W), and performing fermentation experiment for 20h at the early stage and 52h at the later stage. The proportions of the sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp are respectively 1:1:1:2:3:1, 1:2:1:1:1:1, 2:1: 2:3:1, 1:1:2:2:3:1, 1:1:1:2:3:2 and 1:1:1:1:3:1, the total bacterial colony number of the fermentation product, the crude protein (dry basis), the water content (dry basis) and the amino acid (dry basis) are measured, and the results are shown in table 2. As can be seen from table 2, the optimum ratio of sweet potato pulp, tapioca pulp, bran, potato pulp, bean pulp and bean pulp is 2:1:1:2:3: 1.
TABLE 2 influence of different solid media on various physicochemical indices of fermentation products
Example 3
According to the invention, the solid state fermentation time is considered, liquid seed liquid of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and saccharomycetes with the inoculation ratio of 1:1:1:1 is added into a solid culture medium of sweet potato residue, cassava residue, bran, potato residue, bean pulp with the ratio of 2:1:1:2:3:1, the solid culture medium of sweet potato residue, cassava residue, bran, potato residue, bean pulp with the ratio of 2:1:1:2:3:1 is added according to the inoculation amount of 4% (V/W), the fermentation temperature is 37 ℃, and the fermentation time is A: the early stage is aerobic for 0h, and the later stage is half-sealed for 72 h; b: the early stage is aerobic for 10 hours, and the later stage is half-sealed for 62 hours; c: aerobic for 20h in the early stage, and semi-sealing the bag for 52h in the later stage; d: the early stage is aerobic for 36 hours, and the later stage is half-sealed for 36 hours; e: the early stage is aerobic for 52 hours, and the later stage is half-sealed for 20 hours. The total colony count, crude protein (dry basis), moisture (dry basis), amino acids (dry basis) of the fermentation product were determined and the results are shown in Table 3. As shown in Table 3, the optimum solid-state fermentation time was 20 hours in the early stage and 52 hours in the later stage.
TABLE 3 influence of different solid fermentation times on various physicochemical indices of the fermentation product
Example 4
The invention is used for hair-settingThe total colony count, crude protein (dry basis), moisture (dry basis), amino acid (dry basis), lysine, methionine content in the fermentation product were measured and compared to the unfermented material, the results shown in Table 4 indicate that the total colony count in the fermentation product reached 8.2X 1010cfu/g, the crude protein (dry basis) content can reach 24.39%.
TABLE 4 fermentation product index test results
The detection method of the invention comprises the following steps:
determination of colony count: the counts were performed using a dilution plate coating method using an LB solid plate.
Determination of moisture in the fermentation product: the method is characterized by referring to the measurement of the contents of water and other volatile substances in the feed of national standard GB 5009.3-2016.
Determination of amino acids: the amino acids were measured by HPLC.
Determination of crude protein: reference is made to the national standard GB 5009.5-2016.
The technical solutions provided by the embodiments of the present invention are described in detail above, and the principles and implementations of the embodiments of the present invention are described herein by using specific examples, and the descriptions of the above embodiments are only used to help understand the principles of the embodiments of the present invention, and for those skilled in the art, the details and implementations of the embodiments and implementations may be changed, and in summary, the content of the present description should not be understood as a limitation to the present invention.
Claims (10)
1. A preparation method of a fermented feed containing a compound microbial agent is characterized by comprising the following steps:
s1, respectively carrying out liquid fermentation culture on bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast, and then compounding the bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the yeast into a compound microbial agent according to a proportion;
s2, inoculating the compound microbial agent of S1 into a solid culture medium for mixed fermentation; and (3) fermenting at the temperature of 25-45 ℃ for 3-5 days to obtain the fermented feed.
2. The method for preparing the fermented feed containing the compound microbial agent according to claim 1, wherein the bacillus subtilis, the lactobacillus acidophilus, the enterococcus faecium and the yeast are prepared according to the following formula (1-6): (1-6): (1-6): (1-6) in a mass ratio.
3. The method for preparing a fermented feed containing a compound microbial inoculant according to claim 1, wherein the main ingredient in the solid culture medium is at least one of processed sweet potato residue, manioc waste, bran, potato residue, bean dregs and bean pulp.
4. The method for preparing a fermented feed containing a composite microbial inoculant according to claim 2, wherein the ratio of bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast is 1:1:1:1, in a mass ratio of 1.
5. The preparation method of the fermented feed containing the compound microbial agent according to claim 3, wherein the solid culture medium is the processed sweet potato residue, cassava residue, bran, potato residue, bean dregs and bean pulp in a mass ratio of (1-10): (1-10): (1-10): (1-10): (1-10): (1-10) mixing.
6. The method for preparing fermented feed containing compound microbial inoculant according to claim 1, wherein bacillus subtilis, lactobacillus acidophilus, enterococcus faecium and yeast are respectively subjected to liquid fermentation culture, and inoculated into a solid fermentation medium according to the concentration of 2-15% (W/W) for fermentation.
7. The method for preparing a fermented feed containing a complex microbial inoculant according to claim 1, wherein the fermented feed containsThe number of bacterial colonies reaches 1010cfu/g or more.
8. The preparation method of the fermented feed containing the compound microbial agent according to claim 5, wherein the solid culture medium is prepared by mixing the processed sweet potato residue, the cassava residue, the bran, the potato residue, the bean dregs and the bean pulp according to a mass ratio of 2:1:1:2:3: 1.
9. The method for preparing fermented feed containing compound microbial inoculant according to claim 1, wherein in the fermentation, aerobic fermentation is performed for 12-20 h, and then the oxygen concentration is reduced by one time for further fermentation for 52-100 h.
10. The fermented feed containing the compound microbial agent prepared by the preparation method of any one of claims 1 to 9.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210024704.7A CN114304379A (en) | 2022-01-10 | 2022-01-10 | Preparation method of fermented feed containing compound microbial agent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210024704.7A CN114304379A (en) | 2022-01-10 | 2022-01-10 | Preparation method of fermented feed containing compound microbial agent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN114304379A true CN114304379A (en) | 2022-04-12 |
Family
ID=81026120
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210024704.7A Pending CN114304379A (en) | 2022-01-10 | 2022-01-10 | Preparation method of fermented feed containing compound microbial agent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114304379A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115119899A (en) * | 2022-08-30 | 2022-09-30 | 山东泰山生力源集团股份有限公司 | Mixed-strain fermentation microbial preparation and preparation method thereof |
| CN115251234A (en) * | 2022-08-05 | 2022-11-01 | 天津中科新元生物科技有限公司 | Novel process for preparing polypeptide protein feed |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101803682A (en) * | 2009-12-30 | 2010-08-18 | 沈阳科丰牧业科技有限公司 | Compound microbial agent and production method and application thereof |
| CN103027184A (en) * | 2012-12-13 | 2013-04-10 | 浙江德清博诚生物科技有限公司 | Phagostimulant fermented feed using bean pulp, corn protein powder and bran as protein raw materials and a preparation method thereof |
| CN108740312A (en) * | 2018-06-06 | 2018-11-06 | 康源绿洲生物科技（北京）有限公司 | A kind of additive for microbe feedstuff and preparation method thereof |
| CN111066945A (en) * | 2019-12-19 | 2020-04-28 | 广东省微生物研究所（广东省微生物分析检测中心） | Pelletization-free prawn fermented feed |
-
2022
- 2022-01-10 CN CN202210024704.7A patent/CN114304379A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101803682A (en) * | 2009-12-30 | 2010-08-18 | 沈阳科丰牧业科技有限公司 | Compound microbial agent and production method and application thereof |
| CN103027184A (en) * | 2012-12-13 | 2013-04-10 | 浙江德清博诚生物科技有限公司 | Phagostimulant fermented feed using bean pulp, corn protein powder and bran as protein raw materials and a preparation method thereof |
| CN108740312A (en) * | 2018-06-06 | 2018-11-06 | 康源绿洲生物科技（北京）有限公司 | A kind of additive for microbe feedstuff and preparation method thereof |
| CN111066945A (en) * | 2019-12-19 | 2020-04-28 | 广东省微生物研究所（广东省微生物分析检测中心） | Pelletization-free prawn fermented feed |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115251234A (en) * | 2022-08-05 | 2022-11-01 | 天津中科新元生物科技有限公司 | Novel process for preparing polypeptide protein feed |
| CN115119899A (en) * | 2022-08-30 | 2022-09-30 | 山东泰山生力源集团股份有限公司 | Mixed-strain fermentation microbial preparation and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102696860B (en) | Highly efficient and low-cost microbiological feed proteins based on vinegar residue and miscellaneous meal | |
| CN101874546A (en) | Microorganism fermentation feed and preparation method thereof | |
| CN102334611A (en) | Solid-state fermentation method for bacillus natto-saccharomycete composite viable bacteria preparation with rice bran as matrix | |
| CN106173225B (en) | Method for preparing protein feed additive by solid state fermentation of plant protein feed | |
| CN109757605B (en) | Method for producing animal feed by fermenting corn alcohol lees and waste bean dregs | |
| KR20090065428A (en) | A manufacturing method of fermented roughage for ruminants using spent mushroom substrates | |
| CN102234623A (en) | Microbial preparation and fermentation process thereof | |
| CN114304379A (en) | Preparation method of fermented feed containing compound microbial agent | |
| CN113736716B (en) | Lactobacillus paracasei, compound biological leavening agent for yellow storage and yellow storage method | |
| CN114158648A (en) | Tomato peel residue biological fermentation feed, preparation method and application thereof | |
| CN111700157A (en) | Probiotic feed additive for improving immunity of aquatic animals | |
| CN113215051A (en) | Method for preparing feed probiotics by using lactobacillus through rice flour wastewater and passion fruit peel | |
| CN110301526A (en) | Complex micro organism fungicide and its method for preparing bioactive feed | |
| CN107691849B (en) | Feed prepared by microbial fermentation | |
| CN113773996A (en) | Preparation method and application of feeding bacillus subtilis | |
| CN104232547B (en) | It is a kind of for microorganism species additive of sheep feed and preparation method thereof | |
| CN115299527A (en) | Method for degrading vomitoxin in DDGS feed and DDGS feed | |
| CN115316496B (en) | Method for preparing high-protein probiotic animal feed by using waste feathers | |
| CN113875975B (en) | Fermentation process for preparing metaplasia by using wheat processing byproducts | |
| CN113647518B (en) | Special biological fermentation feed for fattening cattle in northern Jiang region and preparation method thereof | |
| CN110973353B (en) | Solid-state fermentation alfalfa meal feed and preparation method and application thereof | |
| CN107897505B (en) | Zymophyte liquid, product containing zymophyte liquid and used for nursing pigs, and preparation method and application of product | |
| CN112136976A (en) | Fermented feed and preparation method thereof | |
| CN112369498A (en) | Composite probiotic preparation for improving low pH condition of rumen of dairy cow and application of composite probiotic preparation in dairy cow feeding | |
| CN110771723A (en) | Cassava residue biological feed and fermentation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20220412 |