CN116711808A - Microbial feed additive and preparation method thereof - Google Patents
Microbial feed additive and preparation method thereof Download PDFInfo
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- CN116711808A CN116711808A CN202310616728.6A CN202310616728A CN116711808A CN 116711808 A CN116711808 A CN 116711808A CN 202310616728 A CN202310616728 A CN 202310616728A CN 116711808 A CN116711808 A CN 116711808A
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- lactobacillus acidophilus
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- 230000000813 microbial effect Effects 0.000 title claims abstract description 68
- 239000003674 animal food additive Substances 0.000 title claims abstract description 50
- 238000002360 preparation method Methods 0.000 title claims abstract description 21
- 238000000855 fermentation Methods 0.000 claims abstract description 95
- 230000004151 fermentation Effects 0.000 claims abstract description 95
- 150000001875 compounds Chemical class 0.000 claims abstract description 11
- 241000193749 Bacillus coagulans Species 0.000 claims description 42
- 244000063299 Bacillus subtilis Species 0.000 claims description 42
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 42
- 240000001046 Lactobacillus acidophilus Species 0.000 claims description 42
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 claims description 42
- 229940054340 bacillus coagulans Drugs 0.000 claims description 42
- 229940039695 lactobacillus acidophilus Drugs 0.000 claims description 42
- 241000235342 Saccharomycetes Species 0.000 claims description 29
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Chemical compound OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 27
- 239000000706 filtrate Substances 0.000 claims description 17
- 238000002156 mixing Methods 0.000 claims description 17
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims description 16
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 16
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 16
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 claims description 14
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- 238000001914 filtration Methods 0.000 claims description 9
- DLRVVLDZNNYCBX-RTPHMHGBSA-N isomaltose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-RTPHMHGBSA-N 0.000 claims description 9
- 229940040461 lipase Drugs 0.000 claims description 9
- 235000019421 lipase Nutrition 0.000 claims description 9
- 235000019319 peptone Nutrition 0.000 claims description 9
- 229940085127 phytase Drugs 0.000 claims description 9
- 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 claims description 8
- BFNBIHQBYMNNAN-UHFFFAOYSA-N ammonium sulfate Chemical compound N.N.OS(O)(=O)=O BFNBIHQBYMNNAN-UHFFFAOYSA-N 0.000 claims description 8
- 229910052921 ammonium sulfate Inorganic materials 0.000 claims description 8
- 235000011130 ammonium sulphate Nutrition 0.000 claims description 8
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 8
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 8
- 238000012216 screening Methods 0.000 claims description 8
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 claims description 7
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- QNAYBMKLOCPYGJ-REOHCLBHSA-N L-alanine Chemical compound C[C@H](N)C(O)=O QNAYBMKLOCPYGJ-REOHCLBHSA-N 0.000 claims description 7
- OUYCCCASQSFEME-QMMMGPOBSA-N L-tyrosine Chemical compound OC(=O)[C@@H](N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-QMMMGPOBSA-N 0.000 claims description 7
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 claims description 7
- AYFVYJQAPQTCCC-UHFFFAOYSA-N Threonine Natural products CC(O)C(N)C(O)=O AYFVYJQAPQTCCC-UHFFFAOYSA-N 0.000 claims description 7
- 239000004473 Threonine Substances 0.000 claims description 7
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 claims description 7
- 235000004279 alanine Nutrition 0.000 claims description 7
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 claims description 7
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 7
- 229960000304 folic acid Drugs 0.000 claims description 7
- 235000019152 folic acid Nutrition 0.000 claims description 7
- 239000011724 folic acid Substances 0.000 claims description 7
- 235000013922 glutamic acid Nutrition 0.000 claims description 7
- 239000004220 glutamic acid Substances 0.000 claims description 7
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 claims description 7
- YZCKVEUIGOORGS-IGMARMGPSA-N Protium Chemical compound [1H] YZCKVEUIGOORGS-IGMARMGPSA-N 0.000 claims description 3
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 3
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- 241000588724 Escherichia coli Species 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 235000015278 beef Nutrition 0.000 description 2
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/23—Lactobacillus acidophilus
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Animal Husbandry (AREA)
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Abstract
The application discloses a microbial feed additive and a preparation method thereof, and belongs to the field of microbial fermentation. Moreover, the optimized culture medium for primary fermentation treatment and the optimized culture medium for secondary fermentation treatment can obviously decompose the catabolite repression effect, are favorable for improving the fermentation of the compound thalli, are used as feed additives, have faint scent, have excellent sense and feeding mouthfeel, and play an excellent role in regulating the balance of intestinal flora of animals.
Description
Technical Field
The application relates to the technical field of microbial fermentation, in particular to a microbial feed additive and a preparation method thereof.
Background
Along with the high-speed development of livestock and poultry cultivation, modern large-scale cultivation is a great trend of livestock and poultry cultivation, and scientific cultivation also has wider application space. However, the mere pursuit of cultivation benefits in the livestock and poultry cultivation industry leads to the potential risks formed for a long time not being properly solved, for example, serious food safety problems are brought about by abuse of antibiotics in feeds, and research on antibiotic substitutes is increasingly paid attention to. In the prior art, the effects of enhancing the organism immunity or muscle enhancement and the like of the livestock and the poultry are achieved by adding traditional Chinese medicine components into the feed, or the microbial balance of the intestinal tracts of the livestock and the poultry is improved by adding a plurality of microbial fermentation agents, so that the feed has the function of replacing antibiotics. However, the existing microbial additions still have the following disadvantages: the interaction between the intestinal microorganisms of livestock and poultry and the environment in a host is a complex and ordered process, in the process, various microorganisms participate in various reactions together, the microbial additive with single microbial agent type can lose activity and function easily due to the change of the environment in the organism, so that the performance is unstable, moreover, the biological activity of the microorganisms with different types and quantity can be exerted only by packing the higher viable count after reaching the intestinal tracts, and the total viable count of the microbial finished product can be reduced along with the influence of factors such as the production technology level, the environment temperature change in the intestinal tracts, hypoxia and the like, so that the use effect of the microbial product is poor. In addition, the intestines and stomach of livestock and poultry are main digestive organs of animals, the microbial products reach the high-bile-salt environment of the intestines through the strong acid environment of the stomach and are deactivated in a large amount, the number of surviving microorganisms is reduced, and after reaching the intestines, the probiotic activity of the microbial products is difficult to be exerted, and the problem of poor use effect of the microbial products is caused. Therefore, it is of great importance to study a highly versatile microbial product.
Disclosure of Invention
Based on the problems, the application provides a microbial feed additive and a preparation method thereof, and the specific technical scheme is as follows:
a microbial feed additive comprising the following preparation raw materials: bacillus coagulans, bacillus subtilis,Yeast and Lactobacillus acidophilus, and each 1kg of the microbial feed additive contains 5.5X10 7 ～6.5×10 7 Bacillus coagulans at a ratio of 2.0X10/g 9 ～3.0×10 9 Bacillus subtilis at a ratio of 1.5X10/g 7 ～2.5×10 7 Yeast of individual/g and 4.0X10 6 ～5.5×10 6 Lactobacillus acidophilus at a concentration of each gram.
Further, the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus are all preserved in a laboratory.
In addition, the preparation method of the microbial feed additive comprises the following steps:
respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
directionally culturing the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state in an acidic environment, mixing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, and performing secondary fermentation treatment to obtain secondary mixed fermentation liquor;
concentrating the secondary mixed fermentation liquor, and drying to obtain the microbial feed additive.
Further, the culture medium for primary fermentation treatment comprises the following components in parts by weight: 1 to 5 parts of isomaltooligosaccharide, 40 to 60 parts of citric acid mycelium pulp, 1 to 3 parts of peptone, 1 to 3 parts of dipotassium hydrogen phosphate and 1 to 3 parts of compound enzyme.
Further, the complex enzyme comprises the following components in percentage by mass (1-3): (7-9): the lipase, cellulase and phytase of (1-2).
Further, the temperature of the primary fermentation treatment is 35-38 ℃, the rotating speed of the remote bed is 150-250 r/min, and the time is 40-48 h.
Further, the pH of the acidic environment is 4.5-6.5.
Further, the secondary fermentation treatment culture medium comprises the following components in parts by weight: 2 to 5 parts of glucose, 3 to 5 parts of xylose, 10 to 15 parts of trehalose, 1 to 3 parts of potassium hydrogen phosphate, 0.1 to 0.5 part of alanine, 0.1 to 0.5 part of glutamic acid, 0.2 to 0.3 part of threonine, 0.1 to 0.3 part of tyrosine, 0.01 to 0.05 part of folic acid, 0.01 to 0.03 part of magnesium sulfate and 0.01 to 0.03 part of ammonium sulfate.
Further, the temperature of the secondary fermentation treatment is 35-38 ℃, the rotating speed of the remote bed is 250-300 r/min, and the time is 28-48 h.
Further, the temperature of the drying treatment is 40-55 ℃.
According to the scheme, the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus are mixed and then subjected to primary fermentation treatment and secondary fermentation treatment, so that the microbial feed additive with excellent comprehensive performance can be obtained after the microbial feed additive is compounded and used, the microbial feed additive is stable in storage and further kept in a stable use state, microbial flora can compete with germs in intestinal tracts of organisms, a host mucosa immune system is enhanced, and the microbial flora can well inhibit colonization and invasion of germs in digestive tracts and further reduce pathogenic risks. In addition, the optimized culture medium for primary fermentation treatment and the optimized culture medium for secondary fermentation treatment can obviously decompose the catabolite repression effect, are favorable for improving the fermentation of the compound thalli, are used as feed additives, have faint scent, have excellent sense and feeding mouthfeel, play a role in excellently regulating the balance of intestinal flora of animals, inhibit the reproduction of escherichia coli and salmonella, obviously improve the growth performance of animals and reduce the pathogenic rate.
Detailed Description
The present application will be described in further detail with reference to the following examples thereof in order to make the objects, technical solutions and advantages of the present application more apparent. It should be understood that the detailed description and specific examples are intended for purposes of illustration only and are not intended to limit the scope of the application.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this application belongs. The terminology used herein in the description of the application is for the purpose of describing particular embodiments only and is not intended to be limiting of the application. The term "and/or" as used herein includes any and all combinations of one or more of the associated listed items.
The application relates to a microbial feed additive and a preparation method thereof, and the specific technical scheme is as follows:
a microbial feed additive comprising the following preparation raw materials: bacillus coagulans, bacillus subtilis, yeast and Lactobacillus acidophilus, and the microbial feed additive was 5.5X10 s per 1kg 7 ～6.5×10 7 Bacillus coagulans at a ratio of 2.0X10/g 9 ～3.0×10 9 Bacillus subtilis at a ratio of 1.5X10/g 7 ～2.5×10 7 Yeast of individual/g and 4.0X10 6 ～5.5×10 6 Lactobacillus acidophilus at a concentration of each gram.
In one embodiment, the bacillus coagulans, the bacillus subtilis, the yeast, and the lactobacillus acidophilus are all stored in a laboratory.
In addition, the preparation method of the microbial feed additive comprises the following steps:
respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
directionally culturing the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state in an acidic environment, mixing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, and performing secondary fermentation treatment to obtain secondary mixed fermentation liquor;
concentrating the secondary mixed fermentation liquor, and drying to obtain the microbial feed additive.
In one embodiment, the primary fermentation medium comprises the following components in parts by weight: 1 to 5 parts of isomaltooligosaccharide, 40 to 60 parts of citric acid mycelium pulp, 1 to 3 parts of peptone, 1 to 3 parts of dipotassium hydrogen phosphate and 1 to 3 parts of compound enzyme.
In one embodiment, the complex enzyme comprises the following components in percentage by mass (1-3): (7-9): the lipase, cellulase and phytase of (1-2). By adding the complex enzyme, the fermentation efficiency and the fermentation effect of the primary fermentation treatment can be promoted.
In one embodiment, the temperature of the primary fermentation treatment is 35-38 ℃, the rotating speed of the remote bed is 150-250 r/min, and the time is 40-48 h.
In one embodiment, the acidic environment has a pH of 4.5 to 6.5.
In one embodiment, the secondary fermentation treatment medium comprises the following components in parts by weight: 2 to 5 parts of glucose, 3 to 5 parts of xylose, 10 to 15 parts of trehalose, 1 to 3 parts of potassium hydrogen phosphate, 0.1 to 0.5 part of alanine, 0.1 to 0.5 part of glutamic acid, 0.2 to 0.3 part of threonine, 0.1 to 0.3 part of tyrosine, 0.01 to 0.05 part of folic acid, 0.01 to 0.03 part of magnesium sulfate and 0.01 to 0.03 part of ammonium sulfate.
In one embodiment, the temperature of the secondary fermentation treatment is 35-38 ℃, the rotating speed of the remote bed is 250-300 r/min, and the time is 28-48 h.
In one embodiment, the temperature of the drying process is 40 ℃ to 55 ℃.
According to the scheme, the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus are mixed and then subjected to primary fermentation treatment and secondary fermentation treatment, so that the microbial feed additive with excellent comprehensive performance can be obtained after the microbial feed additive is compounded and used, the microbial feed additive is stable in storage and further kept in a stable use state, microbial flora can compete with germs in intestinal tracts of organisms, a host mucosa immune system is enhanced, and the microbial flora can well inhibit colonization and invasion of germs in digestive tracts and further reduce pathogenic risks. In addition, the optimized culture medium for primary fermentation treatment and the optimized culture medium for secondary fermentation treatment can obviously decompose the catabolite repression effect, are favorable for improving the fermentation of the compound thalli, are used as feed additives, have faint scent, have excellent sense and feeding mouthfeel, play a role in excellently regulating the balance of intestinal flora of animals, inhibit the reproduction of escherichia coli and salmonella, obviously improve the growth performance of animals and reduce the pathogenic rate.
Embodiments of the present application will be described in detail below with reference to specific examples.
Example 1:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 48 hours under the conditions that the temperature is 38 ℃ and the rotating speed of a remote bed is 150r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 5 parts of isomaltooligosaccharide, 60 parts of citric acid mycelium pulp, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate and the following components in percentage by mass: 7:2, lipase, cellulase and phytase form 3 parts of compound enzyme to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
performing directional culture on the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state under an acidic environment with the pH value of 6.5, mixing the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus, and performing secondary fermentation treatment for 48 hours at the temperature of 37 ℃ and the rotating speed of a remote bed of 250r/min, wherein the culture medium for the secondary fermentation treatment comprises the following components in parts by weight: glucose 5 parts, xylose 5 parts, trehalose 15 parts, potassium hydrogen phosphate 3 parts, alanine 0.5 parts, glutamic acid 0.5 parts, threonine 0.3 parts, tyrosine 0.3 parts, folic acid 0.05 parts, magnesium sulfate 0.03 parts and ammonium sulfate 0.03 parts to obtain a secondary mixed fermentation broth;
concentrating the secondary mixed fermentation liquor, and drying at 45 ℃ to obtain the microbial feed additive.
Example 2:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 45 hours under the conditions that the temperature is 37 ℃ and the rotating speed of a remote bed is 200r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 4 parts of isomaltooligosaccharide, 55 parts of citric acid mycelium pulp, 2 parts of peptone, 2 parts of dipotassium hydrogen phosphate and 3 parts of a weight ratio: 9:1, 2 parts of composite enzyme consisting of lipase, cellulase and phytase to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
performing directional culture on the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state under an acidic environment with the pH value of 6.0, mixing the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus, and performing secondary fermentation treatment for 40 hours at the temperature of 37 ℃ and the rotating speed of a remote bed of 250r/min, wherein the culture medium for the secondary fermentation treatment comprises the following components in parts by weight: 4 parts of glucose, 4 parts of xylose, 12 parts of trehalose, 2 parts of potassium hydrogen phosphate, 0.4 part of alanine, 0.4 part of glutamic acid, 0.2 part of threonine, 0.2 part of tyrosine, 0.03 part of folic acid, 0.02 part of magnesium sulfate and 0.02 part of ammonium sulfate to obtain a secondary mixed fermentation broth;
concentrating the secondary mixed fermentation liquor, and drying at 45 ℃ to obtain the microbial feed additive.
Example 3:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 48 hours under the conditions that the temperature is 36 ℃ and the rotating speed of a remote bed is 150r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 3 parts of isomaltooligosaccharide, 55 parts of citric acid mycelium pulp, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate and 3 parts of starch with the mass ratio of 3:8:1, lipase, cellulase and phytase form 3 parts of compound enzyme to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
performing directional culture on the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state under an acidic environment with the pH value of 6.5, mixing the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus, and performing secondary fermentation treatment for 48 hours at the temperature of 38 ℃ and the distant bed rotation speed of 250r/min, wherein the culture medium for the secondary fermentation treatment comprises the following components in parts by weight: 4 parts of glucose, 4 parts of xylose, 13 parts of trehalose, 2 parts of potassium hydrogen phosphate, 0.4 part of alanine, 0.3 part of glutamic acid, 0.3 part of threonine, 0.2 part of tyrosine, 0.03 part of folic acid, 0.02 part of magnesium sulfate and 0.03 part of ammonium sulfate to obtain a secondary mixed fermentation broth;
concentrating the secondary mixed fermentation liquor, and drying at 50 ℃ to obtain the microbial feed additive.
Comparative example 1:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 48 hours under the conditions that the temperature is 36 ℃ and the rotating speed of a remote bed is 150r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 2 parts of isomaltooligosaccharide, 1 part of xylose, 55 parts of citric acid mycelium pulp, 2 parts of corn starch, 1 part of beef extract, 3 parts of dipotassium hydrogen phosphate and the mass ratio of 3:8:1, lipase, cellulase and phytase form 3 parts of compound enzyme to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
performing directional culture on the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state under an acidic environment with the pH value of 6.5, mixing the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus, and performing secondary fermentation treatment for 48 hours at the temperature of 38 ℃ and the distant bed rotation speed of 250r/min, wherein the culture medium for the secondary fermentation treatment comprises the following components in parts by weight: 4 parts of glucose, 4 parts of xylose, 13 parts of trehalose, 2 parts of potassium hydrogen phosphate, 0.4 part of alanine, 0.3 part of glutamic acid, 0.3 part of threonine, 0.2 part of tyrosine, 0.03 part of folic acid, 0.02 part of magnesium sulfate and 0.03 part of ammonium sulfate to obtain a secondary mixed fermentation broth;
concentrating the secondary mixed fermentation liquor, and drying at 50 ℃ to obtain the microbial feed additive.
Comparative example 2:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 48 hours under the conditions that the temperature is 36 ℃ and the rotating speed of a remote bed is 150r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 3 parts of isomaltooligosaccharide, 55 parts of citric acid mycelium pulp, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate and 3 parts of starch with the mass ratio of 3:8:1, lipase, cellulase and phytase form 3 parts of compound enzyme to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, concentrating the filtrate, and drying at 50 ℃ to obtain the microbial feed additive.
Comparative example 3:
the preparation method of the microbial feed additive comprises the following steps:
each 1kg of the microbial feed additive contains 6.5X10 7 Bacillus coagulans at a ratio of 3.0X10/g 9 Bacillus subtilis at a ratio of 2.5X10/g 7 Yeast of individual/g and 4.0X10 6 Lactobacillus acidophilus at a rate of individual/g; respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment for 48 hours under the conditions that the temperature is 36 ℃ and the rotating speed of a remote bed is 150r/min, wherein the culture medium for the primary fermentation treatment comprises the following components in parts by weight: 3 parts of isomaltooligosaccharide, 55 parts of citric acid mycelium pulp, 3 parts of peptone, 3 parts of dipotassium hydrogen phosphate and 3 parts of starch with the mass ratio of 3:8:1, lipase, cellulase and phytase form 3 parts of compound enzyme to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
performing directional culture on the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state under an acidic environment with the pH value of 6.5, mixing the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus, and performing secondary fermentation treatment for 48 hours at the temperature of 38 ℃ and the distant bed rotation speed of 250r/min, wherein the culture medium for the secondary fermentation treatment comprises the following components in parts by weight: 21 parts of glucose, 10 parts of peptone, 5 parts of beef extract, 0.02 part of magnesium sulfate and 0.03 part of ammonium sulfate to obtain a secondary mixed fermentation broth;
concentrating the secondary mixed fermentation liquor, and drying at 50 ℃ to obtain the microbial feed additive.
The microbial additives obtained in examples 1 to 3 and the microbial additives obtained in comparative examples 1 to 3 were subjected to application tests, and the results are shown in Table 1 below.
Wherein, the test animals are divided into a control group and an experimental group (30 primary lambs in each group), each experimental group is added with 15% of microbial feed additive in basic feed (the experimental group 1 is the microbial additive obtained in the example 1, the experimental group 2 is the microbial additive obtained in the example 2, the experimental group 3 is the microbial additive obtained in the example 3, the comparative experimental group 1 is the microbial additive obtained in the comparative example 1, the comparative experimental group 2 is the microbial additive obtained in the comparative example 2, the comparative experimental group 3 is the microbial additive obtained in the comparative example 3), the lambs are fed with the same basic feed only, and other conditions are kept consistent for 5 weeks. The individual weighing is carried out on the empty stomach experimental group and the control group for unified time every week, and the individual weighing is recorded; after week 5, the lambs of the experimental group and the control group were respectively drenched with 15mL of the mixture with a concentration of 10 10 CFU/mL pathogenic salmonella, lamb morbidity was observed daily.
Table 1:
note that: the same row of data shoulder marks with the same letter indicates that the difference is not significant (P > 0.05), and the shoulder marks with different lower case letters indicate that the difference is significant (P < 0.05).
From the data analysis of Table 1, the microbial feed additive prepared by the application has the effects of promoting the growth of animal organisms and obviously improving daily gain and feed-to-weight ratio.
In addition, the results of statistics on the pathogenicity rate of the lambs in the experimental group and the control group are shown in Table 2 below.
Table 2:
from the data analysis of Table 2, it is known that the incidence of lambs can be limited and reduced by adding the microbial feed additive of the application, and the effect of the microbial additive obtained after fermentation treatment by different processes of the application is also obviously affected, so that the microbial feed additive with obvious effect can be obtained after the optimized fermentation treatment process of the application.
The technical features of the above-described embodiments may be arbitrarily combined, and all possible combinations of the technical features in the above-described embodiments are not described for brevity of description, however, as long as there is no contradiction between the combinations of the technical features, they should be considered as the scope of the description.
The above examples illustrate only a few embodiments of the application, which are described in detail and are not to be construed as limiting the scope of the application. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the application, which are all within the scope of the application. Accordingly, the scope of protection of the present application is to be determined by the appended claims.
Claims (10)
1. The microbial feed additive is characterized by comprising the following preparation raw materials: bacillus coagulans, bacillus subtilis, yeast and Lactobacillus acidophilus, and the microbial feed additive was 5.5X10 s per 1kg 7 ～6.5×10 7 Bacillus coagulans at a ratio of 2.0X10/g 9 ～3.0×10 9 Bacillus subtilis at a ratio of 1.5X10/g 7 ～2.5×10 7 Yeast of individual/g and 4.0X10 6 ～5.5×10 6 Lactobacillus acidophilus at a concentration of each gram.
2. The microbial feed additive according to claim 1, wherein the bacillus coagulans, the bacillus subtilis, the saccharomycetes and the lactobacillus acidophilus are all stored in a laboratory.
3. The preparation method of the microbial feed additive is characterized by comprising the following steps of:
respectively culturing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, mixing and carrying out primary fermentation treatment to obtain primary fermentation liquor;
filtering the primary mixed fermentation liquor, and taking filtrate;
directionally culturing the filtrate, screening out bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus which are excellent in growth state in an acidic environment, mixing bacillus coagulans, bacillus subtilis, saccharomycetes and lactobacillus acidophilus, and performing secondary fermentation treatment to obtain secondary mixed fermentation liquor;
concentrating the secondary mixed fermentation liquor, and drying to obtain the microbial feed additive.
4. The method according to claim 3, wherein the primary fermentation medium comprises the following components in parts by weight: 1 to 5 parts of isomaltooligosaccharide, 40 to 60 parts of citric acid mycelium pulp, 1 to 3 parts of peptone, 1 to 3 parts of dipotassium hydrogen phosphate and 1 to 3 parts of compound enzyme.
5. The method according to claim 4, wherein the complex enzyme comprises the following components in mass ratio (1-3): (7-9): the lipase, cellulase and phytase of (1-2).
6. The method according to claim 4, wherein the primary fermentation treatment is carried out at a temperature of 35-38 ℃, a rotation speed of the remote bed of 150-250 r/min, and a time of 40-48 h.
7. A method of preparation according to claim 3, wherein the pH of the acidic environment is 4.5 to 6.5.
8. A method of preparation according to claim 3, wherein the secondary fermentation treated medium comprises the following components in parts by weight: 2 to 5 parts of glucose, 3 to 5 parts of xylose, 10 to 15 parts of trehalose, 1 to 3 parts of potassium hydrogen phosphate, 0.1 to 0.5 part of alanine, 0.1 to 0.5 part of glutamic acid, 0.2 to 0.3 part of threonine, 0.1 to 0.3 part of tyrosine, 0.01 to 0.05 part of folic acid, 0.01 to 0.03 part of magnesium sulfate and 0.01 to 0.03 part of ammonium sulfate.
9. The preparation method according to claim 8, wherein the temperature of the secondary fermentation treatment is 35-38 ℃, the rotating speed of the remote bed is 250-300 r/min, and the time is 28-48 h.
10. The method according to claim 9, wherein the drying treatment is carried out at a temperature of 40 ℃ to 55 ℃.
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