CN106962609A - A kind of direct putting type biological fermentation feed additive and its production and use - Google Patents
A kind of direct putting type biological fermentation feed additive and its production and use Download PDFInfo
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- CN106962609A CN106962609A CN201710214308.XA CN201710214308A CN106962609A CN 106962609 A CN106962609 A CN 106962609A CN 201710214308 A CN201710214308 A CN 201710214308A CN 106962609 A CN106962609 A CN 106962609A
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Classifications
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- 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
- A23V2400/113—Acidophilus
-
- 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
- A23V2400/125—Casei
-
- 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
- A23V2400/137—Delbrueckii
-
- 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
-
- 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
- A23V2400/51—Bifidobacterium
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- 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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- Fodder In General (AREA)
Abstract
The invention discloses a kind of direct putting type biological fermentation feed additive and its production and use.Feed addictive of the present invention is constituted by auxiliary material, fermentation probiotics, enzyme preparation and with salting liquid culture medium.The product may be directly applied to herding and aquaculture, with raising animal immunizing power, enhancing premunition, improve feed intake and feed conversion rate, treatment and prevention animal intestinal tract disease, the effect for adjusting animal intestinal micro-ecology balance and anti-stress ability.The present invention by production bacterial strain be combined screening, improve purification process and to production bacterial strain production for when carry out strict quality control, solve the problem of biofermentation is unstable.Do not applied directly by any processing after fermentation, it ensure that all bioactive substances produced after fermentation not by any destruction, the problems such as it also avoid transporting simultaneously, store living contaminants, improve effect and security of product, aquaculture cost is considerably reduced, positive impetus is served to green cultivation.
Description
Technical field
It is more particularly to a kind of the present invention relates to a kind of direct putting type biological fermentation feed additive and its production and use
Animal immunizing power, enhancing premunition, raising feed intake and feed conversion rate is improved to play treatment and prevention animal intestinal tract disease, adjust
Save the biological fermentation feed additive of animal intestinal micro-ecology balance and anti-stress ability.Add the invention belongs to biological fermentation feed
Plus agent studying technological domain.
Background technology
Biological fermentation feed has increasingly caused the height of feedstuff industry as a kind of novel fodder in the effect of 21 century
Pay attention to, therefore, biological feedstuff all has very big development prospect in theoretical research or practical application.
Biological feedstuff meets demand of the mankind to health food, the health food of the 21 century mankind will be it is safe and pollution-free,
It is noresidue, rich and varied and possess low fat, low-protein, low in calories, multifilament and have the advantages that prophylactic composition.It is dynamic
The safety of physical property food, is the safety of feed first, therefore, and the safety of feed in itself is to produce the basis of meat product.Biology is raised
Material makes the safety of human food be guaranteed due to the security of its own.
The high speed development of biotechnology promotes the development of biological feedstuff, in the late two decades, and biotechnology has obtained prominent fly
The development pushed ahead vigorously, the separation, purifying, inductive technology to microorganism is more and more perfect, the known microorganisms species with security
More and more, the effect for feed is become better and better, it is ensured that biological feedstuff variety series.
Production biological feedstuff can improve the competitiveness of Feed Enterprise, but this needs the timely application of high-tech technology, together
When be also one of Main Means that Feed Enterprise is contended for markets from now on.Biological feedstuff can effectively improve livestock and poultry and feedstuff is sought
Digesting and assimilating and utilizing for material is supported, the scope of feedstuff application is not only expanded, the problem of feedstuff is nervous is alleviated,
And be more conducive to reduce feed cost, strengthen the competitiveness of Feed Enterprise.The feed of 21 century will be one and be strict with production
Quality, and with cost is low, profit is high, emphasize safe and clean and environmental consciousness feed.Biological feedstuff is due to itself
Characteristic, will reduction nutriment excretion, improve its digestibility and utilization efficiency, save financial resources and manpower in terms of play important
Effect.
Feed can produce a variety of unrighted acids and aromatic acid after microbial fermentation, with special fragranced and good
Good palatability, can obvious stimulation livestock and poultry appetite, increase feed intake.Multiple beneficial bacterium suppresses the life of harmful bacteria in intestines and stomach
Long and breeding, there is special efficacy to intractable diarrhea, and without any antibiotic, no toxicity, side effect.Cholesterol level is low, is
Health care livestock product, meets the mankind to health, health, ecological, environmental protection requirement.Beneficial bacterium can reduce ammonia in pouity dwelling place, vulcanization
The concentration of hydrogen, foul smell, reduces breathing problem, reaches the purpose of purification pouity dwelling place environment, and the immunologic function of stimulating animal, enhancing
Resistance of the animal to a variety of epidemic diseases, hence it is evident that reduction morbidity and mortality.
Biological feedstuff is, as biological feedstuff leavening, feedstuff to be converted into microbial bacteria with microorganism, complex enzyme
The biological fermentation feed that body protein, bioactive micro peptide amino acid, microbial activity probiotics, complex enzyme formulation are integrated.
So-called biological feedstuff is microbiological feed, is that the beneficial microorganism known to mixes warp with feed under microecology theories guidance
Safe and pollution-free, noresidue the high-quality feed containing active probiotic that the special process such as fermentation, dry is made.Biological feedstuff
It is the feed crossed by some special microbial fermentations, and these microorganisms can produce digestive ferment, organic acid, ablastins, B
The materials such as family vitamin, amino acid, by the fermentation to feed, also with regard to benefit materials can be produced, equivalent to prolonging for digestive organs
The increase of long and digestion time, after feed is taken in by animal, various nutritional ingredients can not possibly completely passive thing absorb, do not have
It will be discharged by the composition that animal absorbs with excrement.Animal is higher to the utilization rate of each composition, then the content in excreta is lower,
Pollution to environment is fewer.
Therefore, for environmental protection, nutritionist is simply to carry out some Animal nutrition balances in component design, not
Design, but it is also contemplated that following several respects content in formula design：1. the environmental pollution of stink is controlled；2. improve and control
The environmental pollution of nitrogen (N) processed；3. improve and control the environmental pollution of phosphorus (P)；4. feed digestibility is improved, nutrient is reduced
Loss；5. feed hygiene is improved, the need for development biological feedstuff meets environmental protection, it is known that biological feedstuff can provide the mankind
The safety assurance of food.
Biological feedstuff is widely used in many countries such as European Union, and biological feedstuff also quietly rises in China, China
Biological feedstuff industry development is very fast, and yield increases sharply, and is greatly enriched the market supply of feed, still, and China's biology is raised
Material industry also exist low scale and benefit, technical merit and managerial skills are relatively low, in terms of production cost and labor productivity without competing
The problems such as striving weak advantage, the market competitiveness, the biological feedstuff industrial structure and be laid out unreasonable and biological feedstuff security.
The application present situation of domestic solid biologic feed；(1) Comprehesive biological feed：It is exactly without medicine mixed feed by full nutrition
Add microbial bacterial agent, water and increase the feed through anaerobism or aerobic fermentation at a proper temperature after microbial inoculum mixing.From this
Said in matter, be the new and effective feeding that beneficial active bacteria formulation and perfect compound feed are mixed into fermentation according to a certain percentage and produced
Material.Research on rich enzyme green bio feed shows, can use modern biological project, metabolic control fermentation technology, animal
It is raw that a series of high-tech means such as nutrient balance theory produce the green rich in digestive ferment, lactein and biologically active polypeptide
Thing feed, such feed can not only meet the nutritional need of animal comprehensively, moreover it is possible to increase a variety of digestive ferments, organic acid, function micro-
Biological and polypeptide, the content of amino acid, with the various biological effect such as obvious growth promotion, disease preventing and treating, it is possible to reduce anti-
Raw element, enzyme preparation, the addition of growth promotion health-care agent have preventive and therapeutic action to intestines problem, and can save production cost.
The current country does that not having for solid fermentation complete feed is several, and output is also seldom.(2) non-Comprehesive biological feed：Bioconcentration
Material：The modification feed of fermentation will exactly be coordinated without auxiliary materials such as the concentrated feed of medicine and microorganism and water.Utilize fermentation
The biological feedstuff of agent and supporting technology production can be obviously promoted growth of animal, can especially promote microcirculation and rear buttocks to develop, and carry
High premunition, in substitute antibiotics, in terms of there is uniqueness.
The problem of domestic fermented feed is present；(1) Zymolysis Equipment falls behind, professional lacks；Because most of fermentations are raised
Expect factory, very few to microorganism and zymotechnique understanding, the condition that fermented feed is produced in the sense is all lack of standardization, most
The Zymolysis Equipment of manufacturer and professional are very limited, and some basic conditions just seem particularly poor.Do not meet and defend at all
It is raw to require, microexamination more seriously is carried out without professional at all in them, a reply like this is produced a collection ofly,
So that there is hidden danger in Product Safety.(2) management, sanitary condition are poor；All people for being engaged in microbial fermentation work, which both know about, " to disappear
Poison, sterilizing " work is that one of link is built in the pass of fermentation success or failure.However, many fermented feed factory systems in management are unsound, thing
Stream is chaotic, and employee's health, production environment health all have serious problems.(3) dangerous fermenting raw materials；Many fermented feed producers
Think the raw material typically gone mouldy, as long as ferment with regard to energy detoxification, with regard to that can improve efficiency of feed utilization and improve feed palatability, just
This explanation is understood very little the rudimentary knowledge of " fermentation ".Fermentation is only used for the raw material that fresh, nothing is gone mouldy in principle.In fact make
The plant protein material used in fermented feed is the principal element for having a strong impact on fermented product quality without pollution detection.Many institutes
The serious pathogen of known harm, it is easy to survive and breed in natural environment.They can survive in feedstuff, feed
Long time.Feedstuff is easily salmonella-polluted.Therefore, good storage and feedstuff detection and selection right and wrong
It is often important.(4) strain deactivation prob；It is raw due to the difference of the combination of various microorganism fungus kinds, screening, Cultivated objective and method
Production level, for use the factor difference such as animal species and fermentation processing, condition of storage, Microbe Fermented Soybean Meal production and should
Larger with difference on effect, quality is difficult to ensure that.Beneficial bacterium in Partial fermentation dregs of beans is in culture, dry, packing, preservation, processing
It is dead etc. in production process, the activity of microorganism is reduced, causes product not reach expected effect.Because fermented bean dregs are temporarily without state
Family's standard, each enterprise formulates standard according to the market demand so that in the market fermented bean dregs product fish mesh mixes, and some are even
The microbial fermentation product of personation.(5) strain source is complicated；Because these feed factories are poor to fermentation cognitive ability, it is reluctant to spend
Strain and production technology are bought to regular unit, does not also ask professional to carry out technological guidance, strain security cannot be guaranteed.
Do not did the acute and subacute poison experiment of animal, teratogenesis at all the experiment such as to disable, without product purpose efficacy validation, benefit
Not scientific effect for causing product of raw bacterial content and proportioning is unstable, due to the complexity of animal intestinal tract environment, microorganism
Contained beneficial bacterium number has to reach that certain amount and proportioning just can guarantee that its survival and play a role in feed.Most producers
Understanding to micro-ecology fermentation not enough causes strain improper use, and the product quality produced cannot be stablized and ensure.(6) it is big
Dosage uses growth accelerator；Some feed factories are poor due to fermentation technique level of aggregation, and fermented product miscellaneous bacteria is more, thus by this
When fermented feed is used for concentrated feed for piglet, diarrhea rate is higher, then just increase in concentrated feed for piglet bacitracin, zinc,
The consumption of antibiotic etc..(7) abuse label and mislead user；Some feed factories declare poplar " biological feedstuff " without restraint in packaging bag, " every
Gram contain how many hundred million of viable count " etc., by《Animal microecology》Theory, publicize its effect.
Biological feedstuff has this essential distinction with normal diet：1. by have probiotic microorganisms produce protease, lipase and
Cellulase, can help animal digestion to absorb, and significantly improve the utilization rate of feed.2. palatability is good, and fermented rear feed has
A kind of special fragranced, this is a variety of unrighted acids or aromatic acid produced by microorganism, can substantially irritate pig etc. and move
The appetite of thing.3. multiple beneficial bacterium suppresses the breeding and growth of harmful bacteria in the gastrointestinal tract, prevents from having loose bowels and suffers from diarrhoea.4. without adding
Plus any antibiotic, without any medicament residue, available for production green products of domestic animal.5. biological feedstuff domesticated animal livestock products are used
Product (meat, egg, milk), delicious flavour, the solid ferment content of courage is low, is health care livestock product.6. biological feedstuff nutrition purposes, especially for feeding animals is used, can be made
Ammonia (NH3), the hydrogen sulfide (H of livestock and poultry colony house2S) and the content of scatol is substantially reduced, the purpose of purification barn environment is played.
7. biological feedstuff can be strengthened animal to the premunition of a variety of epidemic diseases, the incidence of disease be greatly lowered with the immunologic function of stimulating animal
And death and culling rate.
In a word, the biological feedstuff of solid fermentation, including Comprehesive biological feed, bioconcentration material, miscellaneous dregs of rice mixed fermentation are formed
New protein source, China rise, have their own characteristics each unlike external, the country is generally using microbe leaven, and state
Naturally occurring lactic acid bacteria class in outer then common use feed；In addition, external, using liquid fermentation feed, the country uses solid
Fermented feed.From the point of view of following development, biological feedstuff may turn into the main flow in market.
In summary, biological feedstuff or fermented feed are a kind of novel fodders, can improve the digestion characteristics of feed, improve dynamic
The production performance of thing, especially effect is obvious in terms of animal feed intake and daily gain, prevention animal diarrhea is improved, it is believed that with hair
The constantly improve of ferment Feed Production Technology, generally will necessarily be received and be widely used by the producer, be conducive to further improving poultry
The level of production is herded, the development of pollution-free food production technology and the competitiveness in international market of enhancing China animal products is promoted.
The content of the invention
The technical problems to be solved by the invention are to propose a kind of direct putting type biological fermentation feed additive and its preparation side
Method and purposes.Biological fermentation feed additive of the present invention is by auxiliary material (dregs of beans, wheat, corn), fermentation probiotics, enzyme
Preparation and constituted with salting liquid culture medium.The product may be directly applied to herding and aquatic farm, with raising animal
Immunity, enhancing premunition, raising feed intake and feed conversion rate, treatment and prevention animal intestinal tract disease, regulation animal intestinal tract
The effect of microecological balance and anti-stress ability.
In order to achieve the above object, present invention employs following technological means：
A kind of direct putting type biological fermentation feed additive of the present invention, it includes following components：
(1) auxiliary material：
Described auxiliary material is made up of dregs of beans, wheat and corn, and wherein the weight ratio of dregs of beans, wheat and corn is 4-8:
0.5-2:1-5, the auxiliary material crosses 60-100 mesh sieves after crushing；
(2) fermentation probiotics
Described fermentation probiotic is according to weight ratio by lactic acid bacteria bacterium powder, bacillus bacterium powder and saccharomycete bacterium powder
80-120:1-10:1-5 is constituted, wherein：
Described lactic acid bacteria bacterium powder is by Bifidobacterium, lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus delbrueckii and cheese
The liquid fermentation liquid of lactobacillus is obtained after frozen drying；
Described bacillus bacterium powder is by one or more kinds of bacillus subtilises and one or more
The liquid fermentation liquid of bacillus licheniformis obtained after frozen drying；
Described saccharomycete bacterium powder is to be freezed by the liquid fermentation liquid of candida tropicalis and saccharomyces cerevisiae through low temperature cold
Obtained after dry；
(3) enzyme preparation
Described enzyme preparation includes neutral proteinase, acid protease and amylase, wherein neutral proteinase：It is acid
Protease：Weight ratio=1 of amylase：1-3：1；
(4) salting liquid culture medium
Described salting liquid culture medium is by 0.1-1.0% (w/w) sodium acetate solution, 0.1-0.5% (w/w) lemon
Sour two ammonium salt solutions, 0.001-0.1% (v/v) solution of Tween 80,0.01-1.0% (w/w) K2HPO4Solution, 0.1-
0.5% (w/w) MgSO4·7H2The MnSO of O solution and 0.001-0.01% (w/w)4·H2O solution is according to isometric mixing
Obtain；
Wherein, the weight ratio of fermentation probiotics, enzyme preparation and salting liquid culture medium is 5-15：1-3：0.5-2；
Wherein, fermentation probiotics, enzyme preparation and the mixture of salting liquid culture medium and the weight ratio of auxiliary material are 1:8-10.
In direct putting type biological fermentation feed additive of the present invention, it is preferred that the weight of dregs of beans, wheat and corn
Amount is than being 6:1:3.
In direct putting type biological fermentation feed additive of the present invention, it is preferred that described fermentation probiotic is by breast
Sour bacterium bacterium powder, bacillus bacterium powder and saccharomycete bacterium powder are 100 according to weight ratio:5:2 compositions.
In direct putting type biological fermentation feed additive of the present invention, it is preferred that described salting liquid culture medium is
By 0.5% (w/w) sodium acetate solution, 0.2% (w/w) dibasic ammonium citrate solution, 0.01% (v/v) Tween 80 is molten
Liquid, 0.1% (w/w) K2HPO4Solution, 0.2% (w/w) MgSO4·7H2The MnSO of O solution and 0.005% (w/w)4·
H2O solution after isometric mixing according to obtaining.
In direct putting type biological fermentation feed additive of the present invention, it is preferred that described Bifidobacterium, acidophilus breast
Bacillus, Lactobacillus plantarum, the deposit number difference CICC 21717 of Lactobacillus delbrueckii and Lactobacillus casei, CICC 6090,
CICC 6073, CICC 20271 and CICC 23486；Described bacillus subtilis is selected from withered with following deposit number
It is more than one or both of careless bacillus：CICC 10453, CICC 10071 and CICC 21095；Described lichens bud
Spore bacillus is selected from more than one or both of bacillus licheniformis with following deposit number：CICC 20684、CICC
21737、CICC 21738；Described candida tropicalis and the deposit number of saccharomyces cerevisiae be respectively CICC32831 and
CICC1832；Above strain is purchased from and is purchased from Chinese industrial Microbiological Culture Collection administrative center (China Center of
Industrial Culture Collection,CICC)。
In direct putting type biological fermentation feed additive of the present invention, it is preferred that fermentation probiotics, enzyme preparation and
The weight ratio of salting liquid culture medium is 10：1.5：1；Ferment probiotics, the mixture of enzyme preparation and salting liquid culture medium with it is auxiliary
The weight ratio of material is 1:9.
In described biological fermentation feed additive of the present invention, the newborn bar of Bifidobacterium, lactobacillus acidophilus, plant
Total Jun Shuo≤10 of bacterium, Lactobacillus delbrueckii and Lactobacillus casei9CFU/g, bacillus subtilis, total bacterium of bacillus licheniformis
Shuo≤2 × 106CFU/g.Total Jun Shuo≤2 × 10 of saccharomycete6CFU/g。
Further, the invention also provides a kind of method for preparing described direct putting type biological fermentation feed additive,
Comprise the following steps：
(1) prepared by auxiliary material：By dregs of beans, wheat, corn according to weight ratio be 4-8:0.5-2:1-5, preferably according to 6:1:3
Crushed after ratio mixing, 60-100 mesh sieves are crossed, by standby after roller drier high temperature drying；
(2) prepared by lactic acid bacteria culturing medium and bacterium powder：
It is prepared by culture medium：Bifidobacterium solid medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, pancreas egg
White peptone 10.0g, dusty yeast 5.0g, glucose 8.0g, Tween 80 1.0g, K2HPO42.0g, sodium acetate 5.0g, hydrogen citrate two
Ammonium 2.0g, ZnSO4·7H2O 0.25g, MgSO4·7H2O 0.1g, agar 15.0g, distilled water are settled to 1L, and pH 6.2~
6.4,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of Bifidobacterium fluid nutrient medium, but it is not added with agar；
Bacillus acidi lactici solid medium is prepared according to following：Casein peptone 10.0g, beef extract 10.0g, dusty yeast 5.0g,
Glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K2HPO42.0g, MgSO4·7H2O
0.2g, MnSO4·H2O 0.05g, CaCO320.0g, agar 15.0g, 1.0L, pH6.8,121 DEG C of 20 minutes high pressures of distilled water are gone out
Bacterium, the same solid medium of preparation of lactic acid bacteria fluid nutrient medium, but it is not added with agar.
Enriched medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone 10.0g, dusty yeast
5.0g, glucose 10.0g, Tween 80 500ml, K2HPO42.0g, sodium acetate 5.0g, distilled water are settled to 1L, and pH 6.2~
6.4,121 DEG C of 20 minutes autoclavings；
It is prepared by lactic acid bacteria bacterium powder:By Bifidobacterium CICC 21717, lactobacillus acidophilus CICC 6090, Lactobacillus plantarum
CICC 6073, Lactobacillus delbrueckii CICC 20271 and Lactobacillus casei CICC 23486 Bifidobacterium solid medium with
And Bifidobacterium, lactobacillus acidophilus, Lactobacillus plantarum, lactic acid are inoculated in respectively after purifying culture on Bacillus acidi lactici solid medium
In galactococcus and Lactobacillus casei fluid nutrient medium, after 37 DEG C of cultures 8 hours by after all strains in equal volume mixing according to connecing
The amount of kind is 1:10 volume ratios are inoculated in the enriched medium in liquid fermentation tank, and after 37 DEG C are cultivated 12 hours, sterile centrifugation is low
Temperature freeze-drying obtains lactic acid bacteria bacterium powder；
(3) prepared by bacillus culture medium and bacterium powder：
It is prepared by bacillus culture medium:Peptone 10.0g, dusty yeast 5.0g, NaCl 10.0g, agar 15.0g, distilled water
1.0L, pH7.0,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of bacillus liquid culture medium, but it is not added with fine jade
Fat；
It is prepared by bacillus bacterium powder：By bacillus subtilis CICC 10453, bacillus subtilis CICC 10071, withered grass
Bacillus CICC 21095, bacillus licheniformis CICC 20684, bacillus licheniformis CICC 21737, bacillus licheniformis
CICC 21738 is inoculated in bacillus liquid culture medium respectively after culture is purified on bacillus solid medium, 37 DEG C
Shaking table 210rpm cultivate 8 hours after by the isometric mixing respectively of bacillus subtilis and bacillus licheniformis, then by withered grass bud
Spore bacillus is with bacillus licheniformis according to volume ratio 1:2 mixing, are then 1 according to inoculum concentration:10 volume ratio is inoculated in liquid
In bacillus liquid culture medium in fermentation tank, after 37 DEG C are cultivated 12 hours, sterile centrifugation, frozen drying obtains gemma
Bacillus bacterium powder；
(4) prepared by microzyme culture medium and bacterium powder：
It is prepared by microzyme culture medium：Dusty yeast 5g, peptone 10g, glucose 20g, agar 15.0g, distilled water 1.0L,
PH6.4,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of saccharomycete fluid nutrient medium, but it is not added with agar；
It is prepared by saccharomycete bacterium powder：Candida tropicalis CICC32831, saccharomyces cerevisiae CICC1832 are trained in saccharomycete solid
Support and be inoculated in respectively in saccharomycete fluid nutrient medium after purifying culture on base, 37 DEG C of shaking table 210rpm cultivate 8 hours after by the torrid zone
It is 1 according to inoculum concentration after Candida is mixed in equal volume with S. cervisiae:10 volume ratio is inoculated in liquid fermentation tank
In saccharomycete fluid nutrient medium, after 37 DEG C are cultivated 12 hours, sterile centrifugation, frozen drying obtains saccharomycete bacterium powder；
(5) preparation of fermentation probiotics：Lactic acid bacteria bacterium powder that above-mentioned steps (2), (3), (4) are obtained, bacillus bacterium
Powder and saccharomycete bacterium powder are according to 80-120:1-10:1-5, preferably according to 100:5:2 part by weight mixing, obtains fermentation benefit
Raw bacterium；
(6) preparation of enzyme preparation：By neutral proteinase, acid protease, amylase according to neutral proteinase：Acidic protein
Enzyme：Amylase=1：1-3：1 part by weight mixing, preferably according to 1：2：1 part by weight mixing；
(7) preparation of salting liquid culture medium：
By 0.1-1.0% (w/w) sodium acetate solution, 0.1-0.5% (w/w) dibasic ammonium citrate solution, 0.001-
0.1% (v/v) solution of Tween 80,0.01-1.0% (w/w) K2HPO4Solution, 0.1-0.5% (w/w) MgSO4·
7H2The MnSO of O solution and 0.001-0.01% (w/w)4·H2O solution according to being mixed to get in equal volume；It is preferred that, it is described
Salting liquid culture medium is by 0.5% (w/w) sodium acetate solution, 0.2% (w/w) dibasic ammonium citrate solution, 0.01% (v/v)
The solution of Tween 80,0.1% (w/w) K2HPO4Solution, 0.2% (w/w) MgSO4·7H2O solution, 0.005% (w/w)
MnSO4·H2O solution according to being mixed to get in equal volume；
(8) salt that the enzyme preparation and step (7) that the fermentation probiotics that obtains step (5), step (6) are obtained are obtained is molten
Liquid culture medium is 5-15 according to weight ratio：1-3：0.5-2 is mixed, and is 10 preferably according to weight ratio：1.5：1 mixing；
(9) auxiliary material that the mixture for obtaining step (8) is obtained with step (1) compares 1 according to weight:8-10, preferably according to
1:9 mixing, as direct putting type biological fermentation feed additive.
In preparation method of the present invention, it is preferred that freeze drying protectant used in described frozen drying
For 0.5% (v/v) glycerite containing 18% (w/w) skimmed milk power and 0.5% (w/w/) lactose.
Further, the invention also provides described direct putting type biological fermentation feed additive is preparing feed addition
Purposes in agent.
Wherein, it is preferred that add its into direct putting type biological fermentation feed additive of the present invention when in use heavy
35-36 DEG C of warm water of 8-10 times of weight is measured, is stirred, is fermented 12 to 18 hours at 25-37 DEG C, it is preferred that also include
1% (w/w) glucose is added into described warm water.
Compared to prior art, the advantage of the invention is that：
(1) it is easy to operate, have a wide range of application, biological fermentation feed additive of the invention is being prepared and supported applied to livestock and poultry
Grow, its feature plant ferments immediately, do not applied directly by any processing after fermentation, it is ensured that all lifes produced after fermentation
Active substances (immune factor, organic acid, antibacterial substance, growth promoting substance, digestive enzymes, bioactive micro peptide and free ammonia
Base is sour and a variety of prebiotics bacterias) not by any destruction, effect of product is greatly improved, aquaculture cost is reduced.
(2) biological feed additive effect of the invention is：Improve immunity；Body is stimulated to produce interferon, promote
Cell division, generation antibody；Strengthen body specificity and nonspecific immune reaction, neutralize endotoxin to strengthen immunity of organism
Ability, improves the resistance to virus.Strengthen premunition；There is antagonism to pathogenic bacteria such as shigella dysenteriae, staphylococcus etc.；Suppression
System and kill the harmful bacterias such as Gram-negative bacteria, Escherichia coli and salmonella, preventing and treating diarrhoea, diarrhea, enteritis, constipation and other
A variety of diseases caused by function of intestinal canal is disorderly, reduction morbidity and the death rate.
(3) feed intake and feed conversion rate are improved：The biological feed additive of the present invention can promote enterogastric peristalsis, significantly carry
High enteron aisle enzyme activity, promotes nutrient digestion to absorb, and improves efficiency of feed utilization.Organic acid produced by probiotics makes calcium, at phosphorus
In the ionic condition easily absorbed, beneficial to skeleton development, promote body growth, reduce animal and occur because of indigestion " feed is just "
Phenomenon, reduce due to immunity inoculation, change feed etc. caused by stress reaction.
(4) ammonia deodorization is dropped：Probiotics can effectively suppress the breeding of the foul smell producing strains such as Escherichia coli substantially to reduce chicken
The odor concentration such as ammonia, hydrogen sulfide in house, reduces the generation of breathing problem and improves breeding environment, lifting cultivation welfare.
(5) bacterial strain combined sorting just to property；We are complete for producing separation, screening, purifying and the inductive technology of bacterial strain
It is kind, and bacterial strain derives from National Industrial DSMZ, with tight security, the effect stability for feed fermentation
Efficiently, biological feedstuff variety series be ensure that.
(6) stable effect：Process operation specification, we employ from the selection of strain and produced the production generation of bacterial strain
Shi Jinhang strict quality controls, it is ensured that the stable effect of biological fermentation feed additive,
(7) security：Biological feed additive is raw materials used and packs by microbial indicators detected, it is to avoid miscellaneous bacteria is dirty
Dye makes biological feed additive ferment effect safety and stability.
(8) aquaculture cost is reduced：The biological fermentation feed of plant produced, drying material biological property reduction increase aquaculture cost
Burden, although biofermentation wet feed biological effect preferably, is not easy to storage, and is easy to pollution microbes, it is straight that we invent
Throwing formula biological fermentation feed additive solves above-mentioned drawback, significantly reduces aquaculture cost, product is served to green cultivation
The impetus of pole.
Embodiment
The invention will now be further described with reference to specific embodiments, advantages of the present invention and feature will be with description and
It is apparent.But these embodiments are only exemplary, do not constitute any limitation to the scope of the present invention.People in the art
Member to the details and form of technical solution of the present invention it should be understood that can enter without departing from the spirit and scope of the invention
Row modifications or substitutions, but these are changed and replacement is each fallen within protection scope of the present invention.
Bacterial strain and its source：
Bifidobacterium (CICC 21717), lactobacillus acidophilus (CICC 6090), Lactobacillus plantarum (CICC 6073), De Shi
Lactobacillus (CICC 20271) and Lactobacillus casei (CICC 23486), bacillus subtilis (CICC 10453), withered grass bud
Spore bacillus (CICC 10071), bacillus subtilis (CICC 21095), bacillus licheniformis (CICC 20684), lichens gemma
Bacillus (CICC 21737), bacillus licheniformis (CICC 21738), candida tropicalis (CICC32831), saccharomyces cerevisiae
(CICC1832) it is purchased from Chinese industrial Microbiological Culture Collection administrative center.
A kind of preparation of direct putting type biological fermentation feed additive of embodiment 1
1st, prepared by auxiliary material：It is 6 by weight afterwards that dregs of beans purchased in market, wheat, corn are qualified by forage standard conventional detection:1:3
Ratio mixing after crush, cross 80 mesh sieves, by standby after (120~150 DEG C) dryings of roller drier high temperature.
2nd, prepared by lactic acid bacteria culturing medium and bacterium powder：
(1) prepared by culture medium：Bifidobacterium solid medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, pancreas
Peptone 10.0g, dusty yeast 5.0g, glucose 8.0g, Tween 80 1.0g, K2HPO42.0g, sodium acetate 5.0g, hydrogen citrate
Two ammonium 2.0g, ZnSO4·7H2O 0.25g, MgSO4·7H2O 0.1g, distilled water is settled to 1L, 6.2~6.4,121 DEG C 20 of pH
Minute autoclaving, the same solid medium of preparation of Bifidobacterium fluid nutrient medium, but it is not added with agar.
Bacillus acidi lactici solid medium is prepared according to following：Casein peptone 10.0g, beef extract 10.0g, dusty yeast 5.0g,
Glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K2HPO42.0g, MgSO4·7H2O
0.2g, MnSO4·H2O 0.05g, CaCO320.0g, agar 15.0g, 1.0L, pH6.8,121 DEG C of 20 minutes high pressures of distilled water are gone out
Bacterium, the same solid medium of preparation of lactic acid bacteria fluid nutrient medium, but it is not added with agar.
Enriched medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone 10.0g, dusty yeast
5.0g, glucose 10.0g, Tween 80 500ml, K2HPO42.0g, sodium acetate 5.0g, distilled water are settled to 1L, and pH 6.2~
6.4,121 DEG C of 20 minutes autoclavings.
(2) prepared by lactic acid bacteria bacterium powder：By Bifidobacterium (CICC 21717), lactobacillus acidophilus (CICC 6090), plant breast
Bacillus (CICC 6073), Lactobacillus delbrueckii (CICC 20271) and Lactobacillus casei (CICC 23486) are solid in Bifidobacterium
Bifidobacterium, lactobacillus acidophilus, plant are inoculated in respectively after purifying culture on body culture medium and Bacillus acidi lactici solid medium
In lactobacillus, Lactococcus lactis and Lactobacillus casei fluid nutrient medium (each bacterium culture medium 400ml), 37 DEG C of cultures 8 are small
When after by the isometric combined inoculation of all strains in (built-in 40 liters of enriched medium of culture medium, connect bacterium in 50 liters of liquid fermentation tanks
Measure as volume ratio 1:10) after 37 DEG C of cultures 12 hours, sterile centrifugation, (- 50 DEG C) of low temperature freeze-drying obtains lactic acid bacteria bacterium powders.
3rd, prepared by bacillus culture medium and bacterium powder：
(1) prepared by bacillus culture medium：Peptone 10.0g, dusty yeast 5.0g, NaCl 10.0g, agar 15.0g, steam
1.0L, pH7.0,121 DEG C of 20 minutes autoclavings of distilled water, the same solid medium of preparation of bacillus liquid culture medium, but not
Plus agar.
(2) prepared by bacillus bacterium powder：By bacillus subtilis (CICC 10453), bacillus subtilis (CICC
10071), bacillus subtilis (CICC 21095), bacillus licheniformis (CICC 20684), bacillus licheniformis (CICC
21737), bacillus licheniformis (CICC 21738) is inoculated in gemma respectively after culture is purified on bacillus solid medium
In bacillus fluid nutrient medium (each bacterium culture medium 200ml), 37 DEG C of shaking table 210rpm cultivate 8 hours after by bacillus subtilis
And bacillus licheniformis isometric mixing respectively, then by bacillus subtilis and bacillus licheniformis in proportion 1:After 2 mixing
It is inoculated in 50 liters of liquid fermentation tanks that (built-in 30 liters of gemma fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1:10), 37 DEG C
After culture 12 hours, sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain bacillus bacterium powder.
4th, prepared by microzyme culture medium and bacterium powder：
(1) prepared by microzyme culture medium：Dusty yeast 5g, peptone 10g, glucose 20g, distilled water 1.0L, pH6.4,121
DEG C 20 minutes autoclavings, the same solid medium of preparation of saccharomycete fluid nutrient medium, but it is not added with agar.
(2) prepared by saccharomycete bacterium powder：By candida tropicalis (CICC32831), saccharomyces cerevisiae (CICC1832) in yeast
It is inoculated in respectively in saccharomycete fluid nutrient medium (each bacterium culture medium 200ml) after purifying culture on bacterium solid medium, 37
DEG C shaking table 210rpm is inoculated in after all strains are mixed in equal volume by candida tropicalis with S. cervisiae after cultivating 8 hours
(built-in 30 liters of saccharomycete fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1 in 50 liters of liquid fermentation tanks:10), 37 DEG C of cultures
After 12 hours, sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain saccharomycete bacterium powder.
5th, prepared by fermentation probiotics：Lactic acid bacteria bacterium powder, bacillus bacterium powder and the yeast that above-mentioned steps 2,3,4 are obtained
Bacterium bacterium powder presses 100:5:2 part by weight mixing is fermentation probiotics.
6th, enzyme preparation：Neutral proteinase (50,000 units/g), acid protease (50,000 units/g), amylase (100,000 units/
G) it is purchase, by neutral proteinase：Acid protease：Amylase=1：2：1 part by weight mixing.
7th, salting liquid culture medium：By 0.5% (w/w) sodium acetate solution, 0.2% (w/w) dibasic ammonium citrate solution,
0.01% (v/v) solution of Tween 80,0.1% (w/w) K2HPO4Solution, 0.2% (w/w) MgSO4·7H2O solution,
0.005% (w/w) MnSO4·H2O solution is mixed in equal volume.
8th, the salting liquid culture that the enzyme preparation and step 7 that the fermentation probiotics that obtains step 5, step 6 are obtained are obtained
Base is 10 according to weight ratio：1.5：1 mixing.
9th, the auxiliary material that the mixture and step 1 obtained step 8 is obtained compares 9 according to weight:1 mixing, as direct putting type are given birth to
Thing feed additive, is dispensed according to every bag of 2kg.
Freeze drying protectant used in being freeze-dried in above-mentioned steps is to contain 18% (w/w) skimmed milk power, 0.5% (w/
W/) 0.5% (v/v) glycerite of lactose.
Animal bifidobacteria, Lactococcus lactis, lactobacillus acidophilus, plant in the biological fermentation feed additive prepared
Total Jun Shuo≤10 of lactobacillus, Lactobacillus casei9CFU/g.Bacillus subtilis, bacillus licheniformis total Jun Shuo≤2 ×
106CFU/g.Total Jun Shuo≤2 × 10 of saccharomycete6CFU/g。
35-36 DEG C of warm water is added in biological fermentation feed additive stir (every bag of 2kg add water 20kg) when using,
25-37 DEG C of fermentation can be used for 12 to 18 hours.
A kind of preparation of direct putting type biological fermentation feed additive of embodiment 2
1st, prepared by auxiliary material：It is 8 by weight afterwards that dregs of beans purchased in market, wheat, corn are qualified by forage standard conventional detection:2:3
Ratio mixing after crush, cross 100 mesh sieves, by standby after (120~150 DEG C) dryings of roller drier high temperature.
2nd, prepared by lactic acid bacteria culturing medium and bacterium powder：
(1) prepared by culture medium：Bifidobacterium solid medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, pancreas
Peptone 10.0g, dusty yeast 5.0g, glucose 8.0g, Tween 80 1.0g, K2HPO4, agar 15.0g, 2.0g, sodium acetate
5.0g, diammonium hydrogen citrate 2.0g, ZnSO4·7H2O 0.25g, MgSO4·7H2O 0.1g, distilled water is settled to 1L, pH 6.2
~6.4,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of Bifidobacterium fluid nutrient medium, but it is not added with agar.
Bacillus acidi lactici solid medium is prepared according to following：Casein peptone 10.0g, beef extract 10.0g, dusty yeast 5.0g,
Glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K2HPO42.0g, MgSO4·7H2O
0.2g, MnSO4·H2O 0.05g, CaCO320.0g, agar 15.0g, 1.0L, pH6.8,121 DEG C of 20 minutes high pressures of distilled water are gone out
Bacterium, the same solid medium of preparation of lactic acid bacteria fluid nutrient medium, but it is not added with agar.
Enriched medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone 10.0g, dusty yeast
5.0g, glucose 10.0g, Tween 80 500ml, K2HPO42.0g, sodium acetate 5.0g, distilled water are settled to 1L, and pH 6.2~
6.4,121 DEG C of 20 minutes autoclavings.
(2) prepared by lactic acid bacteria bacterium powder：By Bifidobacterium (CICC 21717), lactobacillus acidophilus (CICC 6090), plant breast
Bacillus (CICC 6073), Lactobacillus delbrueckii (CICC 20271) and Lactobacillus casei (CICC 23486) are solid in Bifidobacterium
Bifidobacterium, lactobacillus acidophilus, plant are inoculated in respectively after purifying culture on body culture medium and Bacillus acidi lactici solid medium
In lactobacillus, Lactococcus lactis and Lactobacillus casei fluid nutrient medium (each bacterium culture medium 400ml), 37 DEG C of cultures 8 are small
When after by the isometric combined inoculation of all strains in (built-in 40 liters of enriched medium of culture medium, connect bacterium in 50 liters of liquid fermentation tanks
Measure as volume ratio 1:10) after 37 DEG C of cultures 12 hours, sterile centrifugation, (- 50 DEG C) of low temperature freeze-drying obtains lactic acid bacteria bacterium powders.
3rd, prepared by bacillus culture medium and bacterium powder：
(1) prepared by bacillus culture medium：Peptone 10.0g, dusty yeast 5.0g, NaCl 10.0g, agar 15.0g, steam
1.0L, pH7.0,121 DEG C of 20 minutes autoclavings of distilled water, the same solid medium of preparation of bacillus liquid culture medium, but not
Plus agar.
(2) prepared by bacillus bacterium powder：By bacillus subtilis (CICC 10453), bacillus subtilis (CICC
10071), bacillus subtilis (CICC 21095), bacillus licheniformis (CICC 20684), bacillus licheniformis (CICC
21737), bacillus licheniformis (CICC 21738) is inoculated in gemma respectively after culture is purified on bacillus solid medium
In bacillus fluid nutrient medium (each bacterium culture medium 200ml), 37 DEG C of shaking table 210rpm cultivate 8 hours after by bacillus subtilis
And bacillus licheniformis isometric mixing respectively, then by bacillus subtilis and bacillus licheniformis in proportion 1:After 2 mixing
It is inoculated in 50 liters of liquid fermentation tanks that (built-in 30 liters of gemma fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1:10), 37 DEG C
After culture 12 hours, sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain bacillus bacterium powder.
4th, prepared by microzyme culture medium and bacterium powder：
(1) prepared by microzyme culture medium：Dusty yeast 5g, peptone 10g, glucose 20g, distilled water 1.0L, pH6.4,121
DEG C 20 minutes autoclavings, the same solid medium of preparation of saccharomycete fluid nutrient medium, but it is not added with agar.
(2) prepared by saccharomycete bacterium powder：By candida tropicalis (CICC32831), saccharomyces cerevisiae (CICC1832) in yeast
It is inoculated in respectively in saccharomycete fluid nutrient medium (each bacterium culture medium 200ml) after purifying culture on bacterium solid medium, 37
DEG C shaking table 210rpm cultivate 8 hours after by candida tropicalis and S. cervisiae by being inoculated in 50 liters of liquid after mixing in equal volume
(built-in 30 liters of saccharomycete fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1 in fermentation tank:10), 37 DEG C culture 12 hours after,
Sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain saccharomycete bacterium powder.
5th, prepared by fermentation probiotics：Lactic acid bacteria bacterium powder, bacillus bacterium powder and the yeast that above-mentioned steps 2,3,4 are obtained
Bacterium bacterium powder presses 100:3:5 part by weight mixing is fermentation probiotics.
6th, enzyme preparation：Neutral proteinase (50,000 units/g), acid protease (50,000 units/g), amylase (100,000 units/
G) it is purchase, by neutral proteinase：Acid protease：Amylase=1：1：1 part by weight mixing.
7th, salting liquid culture medium：By 0.7% (w/w) sodium acetate solution, 0.5% (w/w) dibasic ammonium citrate solution,
0.05% (v/v) solution of Tween 80,0.05% (w/w) K2HPO4Solution, 0.1% (w/w) MgSO4·7H2O solution,
0.01% (w/w) MnSO4·H2O solution is mixed in equal volume.
8th, the salting liquid culture that the enzyme preparation and step 7 that the fermentation probiotics that obtains step 5, step 6 are obtained are obtained
Base is 8 according to weight ratio：2：1 mixing.
9th, the auxiliary material that the mixture and step 1 obtained step 8 is obtained compares 10 according to weight:1 mixing, as direct putting type are given birth to
Thing feed additive, is dispensed according to every bag of 2kg.
Freeze drying protectant used in being freeze-dried in above-mentioned steps is to contain 18% (w/w) skimmed milk power, 0.5% (w/
W/) 0.5% (v/v) glycerite of lactose.
Animal bifidobacteria, Lactococcus lactis, lactobacillus acidophilus, plant in the biological fermentation feed additive prepared
Total Jun Shuo≤10 of lactobacillus, Lactobacillus casei9CFU/g.Bacillus subtilis, bacillus licheniformis total Jun Shuo≤2 ×
106CFU/g.Total Jun Shuo≤2 × 10 of saccharomycete6CFU/g。
35-36 DEG C of warm water is added in biological fermentation feed additive stir (every bag of 2kg add water 20kg) when using,
25-37 DEG C of fermentation can be used for 12 to 18 hours.
A kind of preparation of direct putting type biological fermentation feed additive of embodiment 3
1st, prepared by auxiliary material：It is 5 by weight afterwards that dregs of beans purchased in market, wheat, corn are qualified by forage standard conventional detection:1:5
Ratio mixing after crush, cross 80 mesh sieves, by standby after (120~150 DEG C) dryings of roller drier high temperature.
2nd, prepared by lactic acid bacteria culturing medium and bacterium powder：
(1) prepared by culture medium：Bifidobacterium solid medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, pancreas
Peptone 10.0g, dusty yeast 5.0g, glucose 8.0g, Tween 80 1.0g, K2HPO4, agar 15.0g, 2.0g, sodium acetate
5.0g, diammonium hydrogen citrate 2.0g, ZnSO4·7H2O 0.25g, MgSO4·7H2O 0.1g, distilled water is settled to 1L, pH 6.2
~6.4,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of Bifidobacterium fluid nutrient medium, but it is not added with agar.
Bacillus acidi lactici solid medium is prepared according to following：Casein peptone 10.0g, beef extract 10.0g, dusty yeast 5.0g,
Glucose 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K2HPO42.0g, MgSO4·7H2O
0.2g, MnSO4·H2O 0.05g, CaCO320.0g, agar 15.0g, 1.0L, pH6.8,121 DEG C of 20 minutes high pressures of distilled water are gone out
Bacterium, the same solid medium of preparation of lactic acid bacteria fluid nutrient medium, but it is not added with agar.
Enriched medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone 10.0g, dusty yeast
5.0g, glucose 10.0g, Tween 80 500ml, K2HPO42.0g, sodium acetate 5.0g, distilled water are settled to 1L, and pH 6.2~
6.4,121 DEG C of 20 minutes autoclavings.
(2) prepared by lactic acid bacteria bacterium powder：By Bifidobacterium (CICC 21717), lactobacillus acidophilus (CICC 6090), plant breast
Bacillus (CICC 6073), Lactobacillus delbrueckii (CICC 20271) and Lactobacillus casei (CICC 23486) are solid in Bifidobacterium
Bifidobacterium, lactobacillus acidophilus, plant are inoculated in respectively after purifying culture on body culture medium and Bacillus acidi lactici solid medium
In lactobacillus, Lactococcus lactis and Lactobacillus casei fluid nutrient medium (each bacterium culture medium 400ml), 37 DEG C of cultures 8 are small
When after by the isometric combined inoculation of all strains in (built-in 40 liters of enriched medium of culture medium, connect bacterium in 50 liters of liquid fermentation tanks
Measure as volume ratio 1:10) after 37 DEG C of cultures 12 hours, sterile centrifugation, (- 50 DEG C) of low temperature freeze-drying obtains lactic acid bacteria bacterium powders.
3rd, prepared by bacillus culture medium and bacterium powder：
(1) prepared by bacillus culture medium：Peptone 10.0g, dusty yeast 5.0g, NaCl 10.0g, agar 15.0g, steam
1.0L, pH7.0,121 DEG C of 20 minutes autoclavings of distilled water, the same solid medium of preparation of bacillus liquid culture medium, but not
Plus agar.
(2) prepared by bacillus bacterium powder：By bacillus subtilis (CICC 10453), bacillus subtilis (CICC
10071), bacillus subtilis (CICC 21095), bacillus licheniformis (CICC 20684), bacillus licheniformis (CICC
21737), bacillus licheniformis (CICC 21738) is inoculated in gemma respectively after culture is purified on bacillus solid medium
In bacillus fluid nutrient medium (each bacterium culture medium 200ml), 37 DEG C of shaking table 210rpm cultivate 8 hours after by bacillus subtilis
And bacillus licheniformis isometric mixing respectively, then by bacillus subtilis and bacillus licheniformis in proportion 1:After 2 mixing
It is inoculated in 50 liters of liquid fermentation tanks that (built-in 30 liters of gemma fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1:10), 37 DEG C
After culture 12 hours, sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain bacillus bacterium powder.
4th, prepared by microzyme culture medium and bacterium powder：
(1) prepared by microzyme culture medium：Dusty yeast 5g, peptone 10g, glucose 20g, distilled water 1.0L, pH6.4,121
DEG C 20 minutes autoclavings, the same solid medium of preparation of saccharomycete fluid nutrient medium, but it is not added with agar.
(2) prepared by saccharomycete bacterium powder：By candida tropicalis (CICC32831), saccharomyces cerevisiae (CICC1832) in yeast
It is inoculated in respectively in saccharomycete fluid nutrient medium (each bacterium culture medium 200ml) after purifying culture on bacterium solid medium, 37
DEG C shaking table 210rpm cultivate 8 hours after by candida tropicalis and S. cervisiae by being inoculated in 50 liters of liquid after mixing in equal volume
(built-in 30 liters of saccharomycete fluid nutrient mediums of culture medium connect bacterium amount for volume ratio 1 in fermentation tank:10), 37 DEG C culture 12 hours after,
Sterile centrifugation, (- 50 DEG C) freeze-dryings of low temperature obtain saccharomycete bacterium powder.
5th, prepared by fermentation probiotics：Lactic acid bacteria bacterium powder, bacillus bacterium powder and the yeast that above-mentioned steps 2,3,4 are obtained
Bacterium bacterium powder presses 100:3:5 part by weight mixing is fermentation probiotics.
6th, enzyme preparation：Neutral proteinase (50,000 units/g), acid protease (50,000 units/g), amylase (100,000 units/
G) it is purchase, by neutral proteinase：Acid protease：Amylase=1：3：1 part by weight mixing.
7th, salting liquid culture medium：By 0.5% (w/w) sodium acetate solution, 0.1% (w/w) dibasic ammonium citrate solution,
0.15% (v/v) solution of Tween 80,0.15% (w/w) K2HPO4Solution, 0.5% (w/w) MgSO4·7H2O solution,
0.005% (w/w) MnSO4·H2O solution is mixed in equal volume.
8th, the salting liquid culture that the enzyme preparation and step 7 that the fermentation probiotics that obtains step 5, step 6 are obtained are obtained
Base is 5 according to weight ratio：2：1 mixing.
9th, the auxiliary material that the mixture and step 1 obtained step 8 is obtained compares 8 according to weight:1 mixing, as direct putting type are given birth to
Thing feed additive, is dispensed according to every bag of 2kg.
Freeze drying protectant used in being freeze-dried in above-mentioned steps is to contain 18% (w/w) skimmed milk power, 0.5% (w/
W/) 0.5% (v/v) glycerite of lactose.
Animal bifidobacteria, Lactococcus lactis, lactobacillus acidophilus, plant in the biological fermentation feed additive prepared
Total Jun Shuo≤10 of lactobacillus, Lactobacillus casei9CFU/g.Bacillus subtilis, bacillus licheniformis total Jun Shuo≤2 ×
106CFU/g.Total Jun Shuo≤2 × 10 of saccharomycete6CFU/g。
35-36 DEG C of warm water is added in biological fermentation feed additive stir (every bag of 2kg add water 20kg) when using,
25-37 DEG C of fermentation can be used for 12 to 18 hours.
Embodiment 4 feeds the biological fermentation feed additive of the present invention to farrowing sow effect assessment
Experiment material：Take 1 bag of direct putting type biological fermentation feed additive (prepared by every bag of 2kg, embodiment 1), plus 36 DEG C of temperature
20 liters of water ((w/w) containing glucose 1%), 37 DEG C of incubators can be used for 12 hours.
Feeding method：30 days before the farrowing sow expected date of childbirth are started with feeding feed additive 100mL, Ran Houtong daily
Cross the effect (table 1) that piglet growing state carrys out reaction additives.
The farrowing sow of table 1 feeds the influence that the biological fermentation feed additive of the present invention grows to piglet daily
Experimental result：The biological fermentation feed additive 100ml that farrowing sow feeds the present invention daily can improve average
Product subnumber 11.7%, average birth weight improves 12.86%, and Piglet at Lactation Period average daily gain improves 14.01%, average weaning weight
12.99% is improved, grice diarrhoea rate declines 72.54%, and the Piglet at Lactation Period death rate declines 78%.
Conclusion：In terms of the experimental result of table 1, farrowing sow feeds this feed additive daily can significantly improve sow
Product subnumber, piglet birth weight, daily gain and weaning weight, significantly reduce grice diarrhoea rate and the death rate.
Embodiment 5 feeds biological fermentation feed additive of the present invention to child care pig effect assessment
Experiment material：Take 1 bag of direct putting type biological fermentation feed additive (prepared by every bag of 2kg, embodiment 1), plus 36 DEG C of temperature
20 liters of water ((w/w) containing glucose 1%) is put into 37 DEG C of incubators and can be used for 12 hours.
Test method：The piglet with a collection of wean is selected, three groups, i.e. additive group, fermented bean dregs group and right is randomly divided into
According to group, each weak sick pig is chosen.Control group ration ingredient is：60% corn flour, concentrate feed 40% (total nitrogen is more than 33%) is real
It is that 5% (w/w) biological fermentation feed additive of the present invention is added in control group feed to test group, and fermented bean dregs group is in control
With the dregs of beans of 5% (w/w) in fermented bean dregs replacement feed in group feed, daily 6 points, 10 points, 14 points, 17 points, at four
Between non-quantitative feed, it is ensured that free choice feeding is not wasted but, and excrement is cleared up daily, record corresponding feed intake, excrement situation (control
Group is conventional to use medicine：Safe glad, florfenicol powder, Qiang Wenxi, auxin etc., which are done, to be prevented, and test group is without medicine), experiment knot
Really (influence of this feed additive to child care pig growth performance is fed daily) be shown in Table 2.
Table 2 feeds influence of this feed additive to child care pig growth performance daily
Experimental result：Child care pig feeds the daily ration containing 5% biological fermentation feed additive of the present invention, average child care daily
Daily gain in pigs experimental group improves 5.23% than control group, improves 4.29% than fermented bean dregs group, feedstuff-meat ratio experimental group compares control group
Reduction by 6.98%, improves 5.26%, diarrhea rate experimental group declines 72.10% than control group, compares fermented bean dregs than fermented bean dregs group
Group declines 64.71%, and three groups of average feed intake differences are not obvious.
Conclusion：In terms of the experimental result of table 2, child care pig feeds this feed additive and can significantly improved daily to increase day by day
Weight, attenuating feedstuff-meat ratio and diarrhea rate.
Feeding this biological fermentation feed Additive On Chickens growth result of embodiment 6 is evaluated
Experiment material：Take 1 bag of direct putting type biological fermentation feed additive (prepared by every bag of 2kg, embodiment 1), plus 36 DEG C of temperature
20 liters of water ((w/w) containing glucose 1%) is put into 37 DEG C of incubators and can be used for 12 hours.
Test method：Selection is randomly divided into 2 groups, i.e. experimental group and control group, control group with 20000 broiler chicken of a collection of selection
The conventional daily ration of feeding, experimental group is that the biological fermentation feed of the present invention of 5% (w/w) of middle addition on the basis of control group daily ration adds
Plus agent, feed 30 days, observe chicken growing state.As a result it is as shown in table 3 below.
Table 3 feeds the influence of this biological fermentation feed Additive On Chickens growth performance
Experimental result：Broiler chicken feeds the daily ration of the biological fermentation feed additive of the present invention of addition 5% (w/w) daily, puts down
Every gross weight improves 2.89% than control group, and feedstuff-meat ratio experimental group is than control group reduction by 4.43%, diarrhea rate experimental group ratio
Control group declines 69.64%, and death rate experimental group is than control group reduction by 44.51%.
Conclusion：In terms of the experimental result of table 3, broiler chicken feeds the biological fermentation feed of the present invention addition of addition 5% (w/w) daily
The daily ration of agent can significantly enhance weight, lower feedstuff-meat ratio, diarrhea rate and the death rate.
In summary, biological fermentation feed additive feature of the invention is to be related to a kind of raising animal immunizing power, enhancing
Premunition, improvement feed conversion rate, which are played, treats and prevents animal intestinal tract disease, adjusts animal intestinal micro-ecology balance and resist and answer
The biological fermentation feed additive for the ability of swashing.The present invention is by being combined screening to production bacterial strain, improving purification technique and right
Produce bacterial strain production for when strict quality control, solve the instability problem of biofermentation.Fermentation directly application in scene ensures
All bioactive substances produced after fermentation are not by any destruction, while it also avoid transporting, storing living contaminants etc. and ask
Topic, improves effect and security of product, considerably reduces aquaculture cost, and positive promotion is served to green cultivation and is made
With.
Claims (10)
1. a kind of direct putting type biological fermentation feed additive, it is characterised in that including following components：
(1) auxiliary material：
Described auxiliary material is made up of dregs of beans, wheat and corn, and wherein the weight ratio of dregs of beans, wheat and corn is 4-8:0.5-
2:1-5, the auxiliary material crosses 60-100 mesh sieves after crushing；
(2) fermentation probiotics
Described fermentation probiotic is 80- according to weight ratio by lactic acid bacteria bacterium powder, bacillus bacterium powder and saccharomycete bacterium powder
120:1-10:1-5 is constituted, wherein：
Described lactic acid bacteria bacterium powder is by the newborn bar of Bifidobacterium, lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus delbrueckii and cheese
The liquid fermentation liquid of bacterium is obtained after frozen drying；
Described bacillus bacterium powder is the ground by one or more kinds of bacillus subtilises and one or more
The liquid fermentation liquid of clothing bacillus is obtained after frozen drying；
Described saccharomycete bacterium powder is the liquid fermentation liquid by candida tropicalis and saccharomyces cerevisiae after frozen drying
Obtain；
(3) enzyme preparation
Described enzyme preparation includes neutral proteinase, acid protease and amylase, wherein neutral proteinase：Acidic protein
Enzyme：Weight ratio=1 of amylase：1-3：1；
(4) salting liquid culture medium
Described salting liquid culture medium is by 0.1-1.0% (w/w) sodium acetate solution, 0.1-0.5% (w/w) citric acid two
Ammonium salt solution, 0.001-0.1% (v/v) solution of Tween 80,0.01-1.0% (w/w) K2HPO4Solution, 0.1-0.5% (w/
W) MgSO4·7H2The MnSO of O solution and 0.001-0.01% (w/w)4·H2O solution according to being mixed to get in equal volume；
Wherein, the weight ratio of fermentation probiotics, enzyme preparation and salting liquid culture medium is 5-15：1-3：0.5-2；
Wherein, fermentation probiotics, enzyme preparation and the mixture of salting liquid culture medium and the weight ratio of auxiliary material are 1:8-10.
2. direct putting type biological fermentation feed additive as claimed in claim 1, it is characterised in that dregs of beans, wheat and corn
Weight ratio is 6:1:3.
3. direct putting type biological fermentation feed additive as claimed in claim 1, it is characterised in that described fermentation probiotic by
Lactic acid bacteria bacterium powder, bacillus bacterium powder and saccharomycete bacterium powder are 100 according to weight ratio:5:2 compositions.
4. direct putting type biological fermentation feed additive as claimed in claim 1, it is characterised in that described salting liquid culture medium
It is that, by 0.5% (w/w) sodium acetate solution, 0.2% (w/w) dibasic ammonium citrate solution, 0.01% (v/v) Tween 80 is molten
Liquid, 0.1% (w/w) K2HPO4Solution, 0.2% (w/w) MgSO4·7H2The MnSO of O solution and 0.005% (w/w)4·
H2O solution after isometric mixing according to obtaining.
5. direct putting type biological fermentation feed additive as claimed in claim 1, it is characterised in that described Bifidobacterium, acidophilus
Lactobacillus, Lactobacillus plantarum, the deposit number difference CICC 21717 of Lactobacillus delbrueckii and Lactobacillus casei, CICC 6090,
CICC 6073, CICC 20271 and CICC 23486；Described bacillus subtilis is selected from withered with following deposit number
It is more than one or both of careless bacillus：CICC 10453, CICC 10071 and CICC 21095；Described lichens bud
Spore bacillus is selected from more than one or both of bacillus licheniformis with following deposit number：CICC 20684、CICC
21737、CICC 21738；Described candida tropicalis and the deposit number of saccharomyces cerevisiae be respectively CICC32831 and
CICC1832；Above strain is purchased from and is purchased from Chinese industrial Microbiological Culture Collection administrative center.
6. direct putting type biological fermentation feed additive as claimed in claim 1, it is characterised in that fermentation probiotics, enzyme preparation with
And the weight ratio of salting liquid culture medium is 10：1.5：1；Ferment probiotics, the mixture of enzyme preparation and salting liquid culture medium with
The weight ratio of auxiliary material is 1:9.
7. a kind of method of the direct putting type biological fermentation feed additive prepared described in claim any one of 1-6, its feature exists
In comprising the following steps：
(1) prepared by auxiliary material：By dregs of beans, wheat, corn according to weight ratio be 4-8:0.5-2:1-5, preferably according to 6:1:3 ratio
Crushed after mixing, 60-100 mesh sieves are crossed, by standby after roller drier high temperature drying；
(2) prepared by lactic acid bacteria culturing medium and bacterium powder：
It is prepared by culture medium：Bifidobacterium solid medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone
10.0g, dusty yeast 5.0g, glucose 8.0g, Tween 80 1.0g, K2HPO42.0g, sodium acetate 5.0g, diammonium hydrogen citrate
2.0g, ZnSO4·7H2O 0.25g, MgSO4·7H2O 0.1g, agar 15.0g, distilled water are settled to 1L, pH 6.2~6.4,
121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of Bifidobacterium fluid nutrient medium, but it is not added with agar；
Bacillus acidi lactici solid medium is prepared according to following：Casein peptone 10.0g, beef extract 10.0g, dusty yeast 5.0g, grape
Sugared 5.0g, sodium acetate 5.0g, dibasic ammonium citrate 2.0g, Tween 80 1.0g, K2HPO42.0g, MgSO4·7H2O 0.2g,
MnSO4·H2O 0.05g, CaCO320.0g, agar 15.0g, 1.0L, pH6.8,121 DEG C of 20 minutes autoclavings of distilled water, breast
The same solid medium of preparation of sour bacteria liquid culture medium, but it is not added with agar.
Enriched medium is prepared according to following：Peptone 5.0g, beef extract 5.0g, tryptone 10.0g, dusty yeast 5.0g, Portugal
Grape sugar 10.0g, Tween 80 500ml, K2HPO42.0g, sodium acetate 5.0g, distilled water are settled to 1L, 6.2~6.4,121 DEG C of pH
20 minutes autoclavings；
It is prepared by lactic acid bacteria bacterium powder；By Bifidobacterium CICC 21717, lactobacillus acidophilus CICC 6090, Lactobacillus plantarum CICC
6073rd, Lactobacillus delbrueckii CICC 20271 and Lactobacillus casei CICC 23486 are in Bifidobacterium solid medium and lactic acid
Bifidobacterium, lactobacillus acidophilus, Lactobacillus plantarum, Lactococcus lactis are inoculated in respectively after purifying culture on bacillus solid medium
And in Lactobacillus casei fluid nutrient medium, according to inoculum concentration be after mixing in equal volume by all strains after 37 DEG C of cultures 8 hours
1:10 volume ratios are inoculated in the enriched medium in liquid fermentation tank, after 37 DEG C are cultivated 12 hours, sterile centrifugation, cryogenic freezing
Dry and obtain lactic acid bacteria bacterium powder；
(3) prepared by bacillus culture medium and bacterium powder：
It is prepared by bacillus culture medium；Peptone 10.0g, dusty yeast 5.0g, NaCl 10.0g, agar 15.0g, distilled water
1.0L, pH7.0,121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of bacillus liquid culture medium, but it is not added with fine jade
Fat；
It is prepared by bacillus bacterium powder：By bacillus subtilis CICC 10453, bacillus subtilis CICC 10071, withered grass gemma
Bacillus CICC 21095, bacillus licheniformis CICC 20684, bacillus licheniformis CICC 21737, bacillus licheniformis CICC
21738 are inoculated in bacillus liquid culture medium respectively on bacillus solid medium after purifying culture, 37 DEG C of shaking tables
210rpm cultivate 8 hours after by the isometric mixing respectively of bacillus subtilis and bacillus licheniformis, then by bacillus subtilis
Bacterium is with bacillus licheniformis according to volume ratio 1:2 mixing, are then 1 according to inoculum concentration:10 volume ratio is inoculated in liquid fermentation
In bacillus liquid culture medium in tank, after 37 DEG C are cultivated 12 hours, sterile centrifugation, frozen drying obtains bacillus
Bacterium powder；
(4) prepared by microzyme culture medium and bacterium powder：
It is prepared by microzyme culture medium：Dusty yeast 5g, peptone 10g, glucose 20g, agar 15.0g, distilled water 1.0L, pH6.4,
121 DEG C of 20 minutes autoclavings, the same solid medium of preparation of saccharomycete fluid nutrient medium, but it is not added with agar；
It is prepared by saccharomycete bacterium powder：By candida tropicalis CICC32831, saccharomyces cerevisiae CICC1832 in saccharomycete solid medium
Be inoculated in respectively in saccharomycete fluid nutrient medium after upper purifying culture, 37 DEG C of shaking table 210rpm cultivate 8 hours after by tropical false silk
It is 1 according to inoculum concentration after yeast is mixed in equal volume with S. cervisiae:10 volume ratio is inoculated in the yeast in liquid fermentation tank
In bacteria liquid culture medium, after 37 DEG C are cultivated 12 hours, sterile centrifugation, frozen drying obtains saccharomycete bacterium powder；
(5) preparation of fermentation probiotics：Lactic acid bacteria bacterium powder that above-mentioned steps (2), (3), (4) are obtained, bacillus bacterium powder with
And saccharomycete bacterium powder is according to 80-120:1-10:1-5, preferably according to 100:5:2 part by weight mixing, obtains probiotics of fermenting；
(6) preparation of enzyme preparation：By neutral proteinase, acid protease, amylase according to neutral proteinase：Acid protease：
Amylase=1：1-3：1 part by weight mixing, preferably according to 1：2：1 part by weight mixing；
(7) preparation of salting liquid culture medium：
By 0.1-1.0% (w/w) sodium acetate solution, 0.1-0.5% (w/w) dibasic ammonium citrate solution, 0.001-0.1%
(v/v) the solution of Tween 80,0.01-1.0% (w/w) K2HPO4Solution, 0.1-0.5% (w/w) MgSO4·7H2O is molten
The MnSO of liquid and 0.001-0.01% (w/w)4·H2O solution according to being mixed to get in equal volume；It is preferred that, described salting liquid
Culture medium be by 0.5% (w/w) sodium acetate solution, 0.2% (w/w) dibasic ammonium citrate solution, 0.01% (v/v's)
The solution of Tween 80,0.1% (w/w) K2HPO4Solution, 0.2% (w/w) MgSO4·7H2O solution and 0.005% (w/
W) MnSO4·H2O solution according to being mixed to get in equal volume；
(8) the salting liquid training that the enzyme preparation and step (7) that the fermentation probiotics that obtains step (5), step (6) are obtained are obtained
It is 5-15 that base, which is supported, according to weight ratio：1-3：0.5-2 is mixed, and is 10 preferably according to weight ratio：1.5：1 mixing；
(9) auxiliary material that the mixture for obtaining step (8) is obtained with step (1) compares 1 according to weight:8-10, preferably according to 1:9 mix
Close, as direct putting type biological fermentation feed additive.
8. preparation method as claimed in claim 7, it is characterised in that frozen-dried protective used in described frozen drying
Agent is 0.5% (v/v) glycerite containing 18% (w/w) skimmed milk power and 0.5% (w/w/) lactose.
9. purposes of the direct putting type biological fermentation feed additive in feed addictive is prepared described in claim 1-6.
10. purposes as claimed in claim 9, it is characterised in that in the direct putting type described in claim any one of 1-6 when using
The 35-36 DEG C of warm water of 8-10 times of weight of its weight is added in biological fermentation feed additive, is stirred, in 25-37 DEG C of fermentation
12 to 18 hours, it is preferred that also include the glucose of from 1% (w/w) of addition to described warm water.
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