CN106387317A - Microorganism feed additive capable of protecting pig livers and preparation method of microorganism feed additive - Google Patents
Microorganism feed additive capable of protecting pig livers and preparation method of microorganism feed additive Download PDFInfo
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- CN106387317A CN106387317A CN201610787368.6A CN201610787368A CN106387317A CN 106387317 A CN106387317 A CN 106387317A CN 201610787368 A CN201610787368 A CN 201610787368A CN 106387317 A CN106387317 A CN 106387317A
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- feed additive
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- saccharomyces cerevisiae
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- 210000004185 liver Anatomy 0.000 title claims abstract description 27
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 244000005700 microbiome Species 0.000 title claims abstract description 14
- 239000003674 animal food additive Substances 0.000 title claims abstract description 10
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 claims abstract description 26
- 238000004519 manufacturing process Methods 0.000 claims abstract description 21
- 108010024636 Glutathione Proteins 0.000 claims abstract description 13
- 229960003180 glutathione Drugs 0.000 claims abstract description 13
- 238000012258 culturing Methods 0.000 claims abstract description 6
- 241000894006 Bacteria Species 0.000 claims description 45
- 238000000855 fermentation Methods 0.000 claims description 36
- 230000004151 fermentation Effects 0.000 claims description 36
- 239000003795 chemical substances by application Substances 0.000 claims description 31
- 240000004808 Saccharomyces cerevisiae Species 0.000 claims description 29
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims description 29
- 239000002131 composite material Substances 0.000 claims description 28
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 claims description 26
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 24
- 239000001963 growth medium Substances 0.000 claims description 24
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 24
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 23
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 23
- 244000063299 Bacillus subtilis Species 0.000 claims description 21
- 239000000047 product Substances 0.000 claims description 21
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 20
- 238000002156 mixing Methods 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 235000013379 molasses Nutrition 0.000 claims description 14
- 229910017053 inorganic salt Inorganic materials 0.000 claims description 13
- 229910052757 nitrogen Inorganic materials 0.000 claims description 13
- 239000004310 lactic acid Substances 0.000 claims description 12
- 235000014655 lactic acid Nutrition 0.000 claims description 12
- 239000000843 powder Substances 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 10
- 239000000654 additive Substances 0.000 claims description 10
- 230000000996 additive effect Effects 0.000 claims description 10
- BVTBRVFYZUCAKH-UHFFFAOYSA-L disodium selenite Chemical compound [Na+].[Na+].[O-][Se]([O-])=O BVTBRVFYZUCAKH-UHFFFAOYSA-L 0.000 claims description 10
- 239000011781 sodium selenite Substances 0.000 claims description 10
- 235000015921 sodium selenite Nutrition 0.000 claims description 10
- 229960001471 sodium selenite Drugs 0.000 claims description 10
- 230000001580 bacterial effect Effects 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 238000012216 screening Methods 0.000 claims description 9
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 8
- 230000001954 sterilising effect Effects 0.000 claims description 8
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 7
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- 229910052799 carbon Inorganic materials 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 7
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- 239000003610 charcoal Substances 0.000 claims description 6
- 238000009423 ventilation Methods 0.000 claims description 6
- 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 5
- 239000001888 Peptone Substances 0.000 claims description 5
- 108010080698 Peptones Proteins 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 5
- 229940041514 candida albicans extract Drugs 0.000 claims description 5
- 239000006052 feed supplement Substances 0.000 claims description 5
- 239000008103 glucose Substances 0.000 claims description 5
- 235000019319 peptone Nutrition 0.000 claims description 5
- 239000011780 sodium chloride Substances 0.000 claims description 5
- 239000012138 yeast extract Substances 0.000 claims description 5
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 claims description 4
- FGUUSXIOTUKUDN-IBGZPJMESA-N C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 Chemical compound C1(=CC=CC=C1)N1C2=C(NC([C@H](C1)NC=1OC(=NN=1)C1=CC=CC=C1)=O)C=CC=C2 FGUUSXIOTUKUDN-IBGZPJMESA-N 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 4
- 235000010469 Glycine max Nutrition 0.000 claims description 4
- 235000013405 beer Nutrition 0.000 claims description 4
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical group [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 4
- 230000036541 health Effects 0.000 claims description 4
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 4
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 4
- 230000002906 microbiologic effect Effects 0.000 claims description 4
- 210000000813 small intestine Anatomy 0.000 claims description 4
- 239000007787 solid Substances 0.000 claims description 4
- 241000186660 Lactobacillus Species 0.000 claims description 3
- GNFTZDOKVXKIBK-UHFFFAOYSA-N 3-(2-methoxyethoxy)benzohydrazide Chemical compound COCCOC1=CC=CC(C(=O)NN)=C1 GNFTZDOKVXKIBK-UHFFFAOYSA-N 0.000 claims description 2
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
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- 235000004977 Brassica sinapistrum Nutrition 0.000 claims description 2
- 210000000582 semen Anatomy 0.000 claims description 2
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- UFHFLCQGNIYNRP-UHFFFAOYSA-N Hydrogen Chemical compound [H][H] UFHFLCQGNIYNRP-UHFFFAOYSA-N 0.000 claims 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 claims 1
- 229910000147 aluminium phosphate Inorganic materials 0.000 claims 1
- 239000011591 potassium Substances 0.000 claims 1
- 229910052700 potassium Inorganic materials 0.000 claims 1
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- 235000020997 lean meat Nutrition 0.000 abstract description 3
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- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 description 2
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- BDKLKNJTMLIAFE-UHFFFAOYSA-N 2-(3-fluorophenyl)-1,3-oxazole-4-carbaldehyde Chemical compound FC1=CC=CC(C=2OC=C(C=O)N=2)=C1 BDKLKNJTMLIAFE-UHFFFAOYSA-N 0.000 description 1
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- ZUBDGKVDJUIMQQ-UBFCDGJISA-N endothelin-1 Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(O)=O)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(O)=O)NC(=O)[C@H]1NC(=O)[C@H](CC=2C=CC=CC=2)NC(=O)[C@@H](CC=2C=CC(O)=CC=2)NC(=O)[C@H](C(C)C)NC(=O)[C@H]2CSSC[C@@H](C(N[C@H](CO)C(=O)N[C@@H](CO)C(=O)N[C@H](CC(C)C)C(=O)N[C@@H](CCSC)C(=O)N[C@H](CC(O)=O)C(=O)N[C@@H](CCCCN)C(=O)N[C@@H](CCC(O)=O)C(=O)N2)=O)NC(=O)[C@@H](CO)NC(=O)[C@H](N)CSSC1)C1=CNC=N1 ZUBDGKVDJUIMQQ-UBFCDGJISA-N 0.000 description 1
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- -1 inorganic acid salt Chemical class 0.000 description 1
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Classifications
-
- 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
- C12N1/18—Baker's yeast; Brewer's yeast
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Organic Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Botany (AREA)
- Fodder In General (AREA)
Abstract
The invention relates to a microorganism feed additive capable of protecting pig livers and a preparation method of the microorganism feed additive. All strains adopted by the microorganism feed additive are screened out from intestinal tracts of animals, and a strain compound culturing method is adopted, so that an antagonism mechanism of the strains is avoided, and fermented products are optimized; and in the whole production technology, beneficial microorganisms are furthest maximized, and a main fermented product namely glutathione is wholly recovered from intracellular and extracellular positions. According to the microorganism feed additive and the preparation method thereof disclosed by the invention, oxygen free radicals can be effectively eliminated, so that the stability of liver cell membranes is improved, the activity of liver enzymes is promoted, the absorption of iron can also be promoted, the integrity of red cell membranes can be maintained, the biosynthesis of DNA is maintained, the normal growth of cells is maintained, and cell immunity is maintained; as a micro-ecologic preparation, the microorganism feed additive can effectively promote the digestion and the absorption of the feed by the animals, the lean meat percentage is increased, toxin disoperation can be alleviated, the feed conversion rate is increased, and the production capacity of bred animals is improved.
Description
Technical field
The present invention relates to feed additive preparing technical field, more particularly, to a kind of microbiological feed of protection pig liver adds
Plus agent and preparation method thereof.
Background technology
Liver is maximum body of gland in pig body, and many materials that hepatocyte produces can directly be released in blood, impact
With the metabolism adjusting body and physiological activity.In addition liver also has conduit system, and the bile of generation can enter 12 fingers through bile duct
Intestinal.Therefore liver has endocrine gland and eccrine property.The blood supply that liver enriches and unique ecologic structure make
Its metabolism is extremely active, not only in terms of the metabolism such as sugar, fat, protein, Metabolism, Vitamins and Hormones with each histoorgan close phase
Close, and there is the critical functions such as secretion, excretion and bioconversion.
However, developing rapidly with scale pig industry, swinery by bad cultivation ideal effect and endangers extremely serious.
Go mouldy the feeding of feedstuff, for a long time in a large number, the use of excess antibiotic, the factor such as the air quality that cultivation density causes greatly is severe,
Being caused directly or indirectly liver, the kidney major injury of pig, thus leading to immunity degradation or even the immunosuppressant of swinery, making pig
It is chronically at sub-health state（" background color disease "）, it is that the invasion of various epidemic diseases provides opportunity, the rustle of leaves in the wind once, pig is
Morbidity.Additionally, the physiological and pathological burden that environmental change stress cause with disease also poses a big pressure to liver.In various pressure
Under, liver is too tired to deal with, and normal function is affected.
Content of the invention
The present invention is directed to defect of the prior art, proposes a kind of microbiological feed promoting barren sow liver oxidation resistance
Additive and preparation method thereof, for realizing this technical purpose, the technical scheme is that：
A kind of preparation method of the additive for microbe feedstuff of protection pig liver is it is characterised in that adopt strain compound criteria side
Method, makes strain tame culture in same culture medium, its preparation process is：
The first step：Bacterial screening and the production production of hybrid seeds
A. bacterial strain screening：Strain source health pig small intestine, carry out lactic acid bacteria using lactic acid bacteria culturing medium MRS and separate, screen plant
Lactobacilluss and bacillus subtilises；Separate saccharomyces cerevisiae with MacConkey agar culture medium in beer fermentation liquid to make in the lab
Bacterial strain preserves；
B. detached Lactobacillus plantarum, bacillus subtilises and saccharomyces cerevisiae are inoculated in broth bouillon respectively, simultaneously
Add the sodium selenite of 0.1-1% ratio, cultivate 20-25 hour at a temperature of 28-35 DEG C, then saccharomyces cerevisiae is inoculated
It is inoculated in nutrient agar in MacConkey agar culture medium, bacillus subtilises, Lactobacillus plantarum is inoculated in the breast in step a
In sour bacterium culture medium, carry out streak culture, select and grow fast, the obvious saccharomyces cerevisiae of thalline feature, Lactobacillus plantarum and hay
Bacillus cereuss, preserve in the lab；
C. produce the production of hybrid seeds：The saccharomyces cerevisiae of step b, Lactobacillus plantarum and bacillus subtilises are transferred respectively into equipped with PDA and
In the Fructus Solani melongenae bottle of MRS nutrient agar, cultivate 40-50 hour at a temperature of 28-35 DEG C, treat Fructus Solani melongenae bottle surface lawn cloth
Full, and middle band can be taken off when yellow in vain, is then placed in being preserved in 2-6 DEG C of refrigerator；
Second step：Composite bacteria agent capable makes
A. weigh each production strain in Fructus Solani melongenae bottle by following weight proportion：Lactobacillus plantarum 5-10%, saccharomyces cerevisiae 70-
85%th, bacillus subtilises 10-20%；
B. strain mixing will be produced, mixed strain will be added on by molasses 2- by the 4-6% of composite bacteria agent capable part by weight
6%th, peptone 0.5-1%, glucose 2-4%, sodium chloride 0-1%, yeast extract 2-6%, magnesium sulfate 0-1%, water 81-93.5% composition
Composite bacteria agent capable culture medium in, adjust ph value to 6-8, put into be heated up in container 110-130 DEG C carry out sterilize 15-20 divide
Clock, in fermentation cylinder for fermentation 6-8 hour after being cooled to 28-35 DEG C, ventilation 10-50L/ hour, mixing speed 180-
250rpm；
3rd step：Composite bacteria agent capable expands and domestication culture
A. composite bacteria agent capable is mixed with charcoal source, nitrogen source, inorganic salt and derivant by following weight proportion：Composite bacteria agent capable 5-15%,
Carbon source 2-15%, nitrogen source 2-10%, inorganic salt 0.5-1%, sodium selenite 0.1-1%, water 58-90.4% mixing insert airtight
In tank, 110-130 DEG C carries out the 15-20 minute that sterilizes, and in fermentation cylinder for fermentation 20-30 hour after being cooled to 28-35 DEG C, leads to
Tolerance 10-50L/ hour, mixing speed 180-250rpm, with the 50% molasses feed supplement sterilizing after 4 hours, 30-60L/ hour,
Until fermentation ends；
B. sample three times in sweat, measure glutathione content, pH value 3.5-4；
4th step：Pulverize and sieve
A. kieselguhr fermentation liquid being added 2% is dried 2-3S at 100-125 DEG C, dried product crosses 80
Mesh sieve, the coarse fodder sifting out is pulverized and is sieved,
B. load in packaging bag after sieving, proceed to warehouse for finished product and stack in order, notify the sampling detection of QC department, major parameter：Water
Part≤12.0%, viable count >=10,000,000,000/g, Glutathione 100mg/100g；Physical behavior：Pale yellow powder shape solid.
Described composite bacteria agent capable is made up of Lactobacillus plantarum, saccharomyces cerevisiae, bacillus subtilises, its weight percent proportioning
For：Lactobacillus plantarum 5-10%, saccharomyces cerevisiae 70-85%, bacillus subtilises 10-20%.
Described carbon source is one of molasses, glucose or their mixture；Described nitrogen source be soybean cake powder, rapeseed cake powder,
One of yeast powder, Semen Maydis pulp or their mixture；Described inorganic salt be one of dipotassium hydrogen phosphate, potassium dihydrogen phosphate or they
Mixture.
On the other hand, the present invention also provides the microbiological feed of the protection pig liver preparing by said method to add
Agent.
The present invention is the strain and organic nutrient substance filtering out in bacterium source using taking food rotten in healthy pig small intestine
Mixing, is made by microbial fermentation technology, it contains various active enzyme, small peptide, free amino acid, monomer
Albumen, antioxidant, vitamin and antibiotic substance, the quantity of probioticss viable count reached 10,000,000,000/gram；
The flora that various kinds of cell of surviving in the intestinal of health pig is constituted, when the beneficial microbe of product of the present invention enters pig road
Afterwards, in conjunction with original probioticss, quickly form superior microorganism flora it is suppressed that the growth of harmful intestinal tract bacteria, therein beneficial
Microorganism is combined closely with intestinal epithelial cell by teichoic acid around intestinal walls, forms Mycoderma barrier, stops harmful bacteria
Field planting；Complex microorganism can also produce antibiotic substance, lactolin, bacitracin etc., suppression escherichia coli, Salmonella etc.
Breeding, field planting and absorption, neutralization toxic product, the synthesis of suppression ammonia and amine in intestinal for the harmful bacteria, promotes the related pouring of intestinal
The highly reactive ability of bar tissue.
In product of the present invention, its contained sulfydryl of contained Glutathione fully can be combined with interior free yl and make
Radical reduction and promote superoxide dismutase synthesis and improve free radical scavenging activity, reductive glutathione can lead to simultaneously
Cross and activate the many kinds of substance metabolism such as the approach such as multiple enzymes promotion saccharide, fat and protein and medicine, bio-toxicity are damaged
Caused renal function injury plays removing toxic substances, stabilizing cell membrane, keeps its integrity and promote the multiple efficacies such as medicine and cellular metabolism.
As a kind of non-peptides endothelin antagonist can effectively antagonism Endothelin, adjust vasomotion, the liver protecting blood vessel endothelium, because
This is to improving liver function, the liver protecting has highly important value.
In product of the present invention, contained active microorganism is beneficial to the utilization rate of absorption raising feedstuff and the energy of nutrient, fall
Low feed-weight ratio；Additional inorganic acid salt can be reduced, reduce the discharge of feces phosphorus, mitigate the pollution to environment, but also can improve
Combined protein, the utilization of mineral element, improve digestibility.
Product of the present invention adopts carbon source, nitrogen source, inorganic salt and sodium selenite as spawn culture raw material, and they are microorganisms
Cellularity and the important component part of enzyme, also can maintain the activity of enzyme, can promote microbial growth, stimulating activity enzyme and
The generation of small peptide.
The purpose that the present invention cultivates from ecological ideas is started with, detached probiotics strain in selection-breeding nature, useless with agricultural
Material raw material is culture medium, by multistage enrichment fermentation, produces being beneficial to Multiple components digestion and suction in feedstuff of high concentration
The profitable strain received, produces the additive of the material such as Glutathione and small peptide simultaneously, directly adds, can improve pig in feedstuff
Liver antioxidant activity, can be by promoting the secretion of hypophysis GH, and GH may be by raising liver GHR gene transcription level and GHR
Level, strengthens liver, semitendinosus m. IGF-1 genetic transcription, reduces adipose cell IGF-1 genetic transcription, so that serum I GF-1 level is carried
Height, and then promote albumen synthesis, improve plant recovery of nutrient, the final growth promoting pig.Liver can purify by animal absorb mould
Verticillium toxin, the redox reaction based on Glutathione for this purification process, toxin infringement can be mitigated.The present invention is a kind of peace
Entirely, efficient green feed additive.
In sum, the present invention with respect to prior art advantage be：
1., using compound criteria and the fermentation technique that combines of domestication of original creation, it is to avoid the Antagonizing of each strain, make fermentation
Product obtains optimization；
2. whole production technology obtains beneficial microbe maximization to the full extent, main product Glutathione intracellular born of the same parents of fermenting
All reclaim outward；
3. in product, contain the lactic acid promoting feedstuff feed intake, the cellulase producing in also fermenting, protease, Digestive Enzyme etc.
Various active enzyme, with profitable strain collective effect, makes the nutrition in feedstuff fully absorb utilization.
4. in the present invention, main raw material is agricultural by-products molasses, along with using cereal crops and common feedstuffses carrier
As main medium raw material, low production cost is it is easy to promote in cultivation.
5. the product of the present invention adopts 2045 in feed industry（2013）The strain of bulletin is fermented, and is ensureing to improve
Pig self immune system and growth promoter, moreover it is possible to accelerate antibiotic, heavy metal and toxin metabolism, meet the requirement of the ecological agriculture,
Safe green product is provided to provide safeguard for the people.
Specific embodiment
The invention provides a kind of preparation method of the additive for microbe feedstuff of protection pig liver, comprise the following steps：
Step a. bacterial strain screening：Strain source health pig small intestine, carry out lactic acid bacteria using lactic acid bacteria culturing medium and separate, screening is planted
Thing lactobacilluss and bacillus subtilises；MacConkey agar culture medium is used to separate saccharomyces cerevisiae in beer fermentation liquid；
Detached Lactobacillus plantarum, bacillus subtilises and saccharomyces cerevisiae are inoculated in broth bouillon step b. respectively,
Add the sodium selenite of 0.1-1% ratio simultaneously, cultivate 20-25 hour at a temperature of 28-35 DEG C, then by saccharomyces cerevisiae
It is inoculated in MacConkey agar culture medium, bacillus subtilises are inoculated in nutrient agar, Lactobacillus plantarum is inoculated in lactic acid bacteria
In culture medium, carry out streak culture, select and grow fast, the obvious saccharomyces cerevisiae of thalline feature, Lactobacillus plantarum and hay spore
Bacillus, preserves；
Step c. produces the production of hybrid seeds：The saccharomyces cerevisiae obtaining in step b, Lactobacillus plantarum and bacillus subtilises are transferred respectively
Enter in the culture bottle equipped with PDA and MRS nutrient agar, cultivate 40-50 hour at a temperature of 28-35 DEG C, wait to cultivate
Bottle surface lawn is covered with, and middle band can be taken off when yellow in vain, is then placed in being saved backup in 2-6 DEG C of refrigerator；
Step d. weighs each production strain saving backup in culture bottle in step c by weight ratio, then mixes three；
Mixed production strain is added on composite bacteria agent capable culture medium by the 4-6% of composite bacteria agent capable part by weight by step e.
In, adjust ph value to 6-8, put into be heated up in container 110-130 DEG C carry out sterilize 15-20 minute, be cooled to 28-35
In fermentation cylinder for fermentation 6-8 hour after DEG C, obtain composite bacteria agent capable；
The composite bacteria agent capable that step e obtains is mixed by step f. with charcoal source, nitrogen source, inorganic salt, derivant and water, 110-130 DEG C
Carry out the 15-20 minute that sterilizes, in fermentation cylinder for fermentation 20-30 hour after being cooled to 28-35 DEG C, ventilation 10-50L/ hour,
Mixing speed 180-250rpm, with the 50% molasses feed supplement sterilizing after 4 hours, 30-60L/ hour, until fermentation ends；
The kieselguhr that fermentation liquid adds 2% is dried 2-3S, dried product mistake at 100-125 DEG C by step g.
80 mesh sieves, the coarse fodder sifting out is pulverized and is sieved, and obtains the additive for microbe feedstuff of protection pig liver.
Below by specific embodiment, the present invention is described in more detail, for a better understanding of the present invention,
But following embodiments are not intended to limit the scope of the invention.
Embodiment 1
The first step：Bacterial screening and the production production of hybrid seeds
A. bacterial strain screening：Taking food rotten in healthy goose small intestinal is bacterium source, carries out using by the lactic acid bacteria culturing medium of MRS culture medium
Lactic acid bacteria separates, screens Lactobacillus plantarum and bacillus subtilises；Use PDA agar culture medium to separate in beer fermentation liquid to make wine
Yeast is made bacterial strain in the lab and is preserved, and the present invention uses following strain：Lactobacillus plantarum Lactobacillus
plantarum（ATCC8014）, Saccharomyces Cerevisiae in S accharomyces cerevisiae（IFO0203）, bacillus subtilises
Bacillus subtilis （ACCC10623）；
B. detached Lactobacillus plantarum, bacillus subtilises and saccharomyces cerevisiae are inoculated in broth bouillon respectively, simultaneously
Add the sodium selenite of 0.5% ratio, cultivate 24 hours at a temperature of 30 DEG C, then saccharomyces cerevisiae is inoculated in by glucose
1g, potassium chloride 1.8g, yeast extract 2.5g, Sodium Acetate Trihydrate 8.2g, the Maxwell of agar 15-20g, distilled water 1000ml composition
（Meclary）In agar culture medium, bacillus subtilises are inoculated in by Carnis Bovis seu Bubali cream 3g, peptone 10g, sodium chloride 5g, agar
15-20g, the Nutrient agar of water 1000ml, pH7.0-7.2 composition（LB culture medium）, Lactobacillus plantarum is inoculated in step a
In lactic acid bacteria culturing medium, carry out streak culture, select that growth is fast, the obvious saccharomyces cerevisiae of thalline feature, Lactobacillus plantarum and withered
Careless bacillus cereuss, preserve in the lab；
C. produce the production of hybrid seeds：The saccharomyces cerevisiae of step b, Lactobacillus plantarum and bacillus subtilises are transferred respectively into equipped with nutrition
In the Fructus Solani melongenae bottle of agar culture medium, cultivate 45 hours at a temperature of 30 DEG C, treat that Fructus Solani melongenae bottle surface lawn is covered with, and when middle band is yellow in vain
Can be taken off;Preserved in the refrigerator being then placed in 4 DEG C;
Above isolation medium all can be found in Laboratory Manual.
Second step：Composite bacteria agent capable makes
A. weigh each production strain in Fructus Solani melongenae bottle by following weight proportion：Lactobacillus plantarum 10%, saccharomyces cerevisiae 80%, withered
Careless bacillus cereuss 10%.
B. strain mixing will be produced, be added on by molasses 4%, peptone 0.8%, Fructus Vitis viniferae in 4% ratio of culture medium weight
In sugar 3%, sodium chloride 0.5%, yeast extract 2%, magnesium sulfate 0.05%, the composite bacteria agent capable culture medium of water 89.65% composition, ph value is adjusted
To 7.0, put into and be heated up to 120 DEG C in container and carry out sterilizing 15 minutes, be cooled in 30 DEG C of after fermentation tanks and ferment 8 hours, ventilation
Amount 20L/ hour, mixing speed 200rpm；
3rd step：Composite bacteria agent capable expands and domestication culture
A. composite bacteria agent capable is mixed with charcoal source, nitrogen source, inorganic salt and derivant by weight ratio：Composite bacteria agent capable
10% transfers into containing molasses 4%（Carbon source）, soybean cake powder 2%（Nitrogen source）, dipotassium hydrogen phosphate 0.4, potassium dihydrogen phosphate 0.2%
（Inorganic salt）, sodium selenite 0.5%, water 92.9% mixing insert in vapor tight tank, be heated up to 120 DEG C and carry out sterilizing 15 minutes, cold
But to after 30 DEG C in fermentation cylinder for fermentation 24 hours, ventilation 40L/ hour, mixing speed 200rpm, with 50% sterilizing after 4 hours
The molasses feed supplement crossed, 36L/ hour, until fermentation ends；
B. sample three times in sweat, measure glutathione content, pH value 3.5-4.
4th step：Pulverize and sieve
A. kieselguhr fermentation liquid being added 2% is dried 2.5S at 115 DEG C, dried product crosses 80 mesh sieves,
The coarse fodder sifting out is pulverized and is sieved,
B. load in packaging bag after sieving, proceed to warehouse for finished product and stack in order, notify the sampling detection of QC department, major parameter：Water
Part≤12.0%, viable count >=10,000,000,000/g, Glutathione 100mg/100g；Physical behavior：Pale yellow powder shape solid.
Embodiment 2
The first step:Bacterial screening is identical with embodiment 1 with the production production of hybrid seeds；
Second step：Composite bacteria agent capable makes
A. weigh each production strain in Fructus Solani melongenae bottle by following weight proportion：Lactobacillus plantarum 8%, saccharomyces cerevisiae 80%, hay
Bacillus cereuss 12%.
B. strain mixing will be produced, be added on by molasses 5%, peptone 0.6%, Fructus Vitis viniferae in 4% ratio of culture medium weight
In sugar 3%, sodium chloride 0.8%, yeast extract 2.5%, magnesium sulfate 0.08%, the composite bacteria agent capable culture medium of water 88.02% composition, ph value is adjusted
Save to 7.0, put into and be heated up to 121 DEG C in container and carry out sterilizing 15 minutes, be cooled in 30 DEG C of after fermentation tanks and ferment 8 hours, lead to
Tolerance 25L/ hour, mixing speed 180rpm；
3rd step：Composite bacteria agent capable expands and domestication culture
A. composite bacteria agent capable is mixed with charcoal source, nitrogen source, inorganic salt and derivant by weight ratio：Composite bacteria agent capable
15% transfers into containing molasses 5%（Carbon source）, soybean cake powder 2.5%（Nitrogen source）, dipotassium hydrogen phosphate 0.5, potassium dihydrogen phosphate
0.25%（Inorganic salt）, sodium selenite 0.8%, water 90.95% mixing insert in vapor tight tank, be heated up to 121 DEG C and carry out sterilizing 15
Minute, in fermentation cylinder for fermentation 26 hours after being cooled to 30 DEG C, ventilation 30L/ hour, mixing speed 220rpm, use after 4 hours
The molasses feed supplement that 50% sterilized, 30L/ hour, until fermentation ends；
B. sample three times in sweat, measure glutathione content, pH value 3.5-4.
4th step：Pulverize and sieve
A. kieselguhr fermentation liquid being added 2% is dried 2.5S at 118 DEG C, dried product crosses 80 mesh sieves,
The coarse fodder sifting out is pulverized and is sieved,
B. load in packaging bag after sieving, proceed to warehouse for finished product and stack in order, notify the sampling detection of QC department, major parameter：Water
Part≤12.0%, viable count >=10,000,000,000/g, Glutathione 100mg/100g；Physical behavior：Pale yellow powder shape solid.
It is added to obtaining in embodiment 1 and 2 in pig feed in 200-400ppm ratio when additive uses and stir all
Even, then measure index of correlation.This product will substantially be increased using daily pig food ration in feedstuff, be SOD in test sera
Content improves 20%, MDA than comparison and have dropped more than 10%, feedstuff research and development centre of Shanghai academy of agricultural sciences carry out to having a competition
In testing, add the product of 200ppm present invention preparation, fatten daily gain in pigs and be respectively increased 11.67%；Feed conversion rate improves
8.9%；Carcass lean meat percentage improves 5.80%.It can be seen that, add this product in finishing pigs diets in growth promotion, raising lean meat percentage, saving
The aspect effect such as feedstuff shows.
Above the specific embodiment of the present invention is described in detail, but it has been intended only as example, the present invention has not limited
It is formed on particular embodiments described above.To those skilled in the art, any equivalent modifications that the present invention is carried out and
Substitute also all among scope of the invention.Therefore, the impartial conversion made without departing from the spirit and scope of the invention and
Modification, all should cover within the scope of the invention.
Claims (10)
1. a kind of preparation method of the additive for microbe feedstuff of protection pig liver is it is characterised in that comprise the following steps：
Step a. bacterial strain screening：Strain source health pig small intestine, carry out lactic acid bacteria using lactic acid bacteria culturing medium and separate, screening is planted
Thing lactobacilluss and bacillus subtilises；MacConkey agar culture medium is used to separate saccharomyces cerevisiae in beer fermentation liquid；
Detached Lactobacillus plantarum, bacillus subtilises and saccharomyces cerevisiae are inoculated in broth bouillon step b. respectively,
Add the sodium selenite of 0.1-1% ratio simultaneously, cultivate 20-25 hour at a temperature of 28-35 DEG C, then by saccharomyces cerevisiae
It is inoculated in MacConkey agar culture medium, bacillus subtilises are inoculated in nutrient agar, Lactobacillus plantarum is inoculated in lactic acid bacteria
In culture medium, carry out streak culture, select and grow fast, the obvious saccharomyces cerevisiae of thalline feature, Lactobacillus plantarum and hay spore
Bacillus, preserves；
Step c. produces the production of hybrid seeds：The saccharomyces cerevisiae obtaining in step b, Lactobacillus plantarum and bacillus subtilises are transferred respectively
Enter in the culture bottle equipped with PDA and MRS nutrient agar, cultivate 40-50 hour at a temperature of 28-35 DEG C, wait to cultivate
Bottle surface lawn is covered with, and middle band can be taken off when yellow in vain, is then placed in being saved backup in 2-6 DEG C of refrigerator；
Step d. weighs each production strain saving backup in culture bottle in step c by weight ratio, then mixes three；
Mixed production strain is added on composite bacteria agent capable culture medium by the 4-6% of composite bacteria agent capable part by weight by step e.
In, adjust ph value to 6-8, put into be heated up in container 110-130 DEG C carry out sterilize 15-20 minute, be cooled to 28-35
In fermentation cylinder for fermentation 6-8 hour after DEG C, obtain composite bacteria agent capable；
The composite bacteria agent capable that step e obtains is mixed by step f. with charcoal source, nitrogen source, inorganic salt, derivant and water, 110-130 DEG C
Carry out the 15-20 minute that sterilizes, in fermentation cylinder for fermentation 20-30 hour after being cooled to 28-35 DEG C, ventilation 10-50L/ hour,
Mixing speed 180-250rpm, with the 50% molasses feed supplement sterilizing after 4 hours, 30-60L/ hour, until fermentation ends；
The kieselguhr that fermentation liquid adds 2% is dried 2-3S, dried product mistake at 100-125 DEG C by step g.
80 mesh sieves, the coarse fodder sifting out is pulverized and is sieved, and obtains the additive for microbe feedstuff of protection pig liver.
2. preparation method according to claim 1 is it is characterised in that respectively produce the weight percent of strain in described step d
Proportioning is：Lactobacillus plantarum 5-10%, saccharomyces cerevisiae 70-85%, bacillus subtilises 10-20%.
3. preparation method according to claim 1 is it is characterised in that described composite bacteria agent capable culture medium is according to weight proportion group
It is divided into molasses 2-6%, peptone 0.5-1%, glucose 2-4%, sodium chloride 0-1%, yeast extract 2-6%, magnesium sulfate 0-1%, water 81-
93.5%.
4. preparation method according to claim 1 is it is characterised in that derivant described in step f is sodium selenite.
5. preparation method according to claim 4 it is characterised in that composite bacteria agent capable and charcoal source in described step f, nitrogen source,
Inorganic salt, derivant and water mixing part by weight be：5-15% composite bacteria agent capable, carbon source 2-15%, nitrogen source 2-10%, inorganic salt
0.5-1%, sodium selenite 0.1-1%, water 58-90.4%.
6. preparation method according to claim 1 is it is characterised in that described carbon source is selected from molasses or glucose at least
A kind of.
7. preparation method according to claim 1 is it is characterised in that described nitrogen source is selected from soybean cake powder, rapeseed cake powder, yeast
At least one in powder and Semen Maydis pulp.
8. preparation method according to claim 1 is it is characterised in that above-mentioned inorganic salt is selected from dipotassium hydrogen phosphate, phosphoric acid
At least one of potassium dihydrogen.
9. the microbiological feed of the protection pig liver that preparation method according to claim 1-8 any one prepares adds
Agent.
10. the additive for microbe feedstuff of protection pig liver according to claim 9 is it is characterised in that described microorganism
Feed additive is pale yellow powder shape solid, moisture content≤12.0%, viable count >=10,000,000,000/g, Glutathione >=100mg/
100g.
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| CN108112786A (en) * | 2017-12-18 | 2018-06-05 | 中国农业大学 | Application of the cysteine as enteron aisle essential nutrients in animal health maintenance |
| CN108522841A (en) * | 2018-03-19 | 2018-09-14 | 牡丹江市艾利维亚生物科技有限公司 | A kind of animals and plants nanometer selenium peptide nutrient solution and preparation method thereof |
| CN116998593A (en) * | 2023-09-15 | 2023-11-07 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Application of lactobacillus plantarum in preparation for improving fish sugar tolerance |
| CN117904009A (en) * | 2024-03-19 | 2024-04-19 | 深圳中科翎碳生物科技有限公司 | Bacillus subtilis applicable to non-grain bio-based carbon source and fermentation production method thereof |
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| CN106858040A (en) * | 2017-03-15 | 2017-06-20 | 湖北绿科乐华生物科技有限公司 | High content reduced glutathione Cultures of S. cerevisiae and its production method |
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| CN117904009A (en) * | 2024-03-19 | 2024-04-19 | 深圳中科翎碳生物科技有限公司 | Bacillus subtilis applicable to non-grain bio-based carbon source and fermentation production method thereof |
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