CN110214871A - A kind of layer breeding probiotics and preparation method thereof - Google Patents
A kind of layer breeding probiotics and preparation method thereof Download PDFInfo
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- CN110214871A CN110214871A CN201910354362.3A CN201910354362A CN110214871A CN 110214871 A CN110214871 A CN 110214871A CN 201910354362 A CN201910354362 A CN 201910354362A CN 110214871 A CN110214871 A CN 110214871A
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Classifications
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- A—HUMAN NECESSITIES
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- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
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- A23K10/18—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions of live microorganisms
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- A—HUMAN NECESSITIES
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- C—CHEMISTRY; METALLURGY
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- C—CHEMISTRY; METALLURGY
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- 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
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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
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- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The present invention provides a kind of layer breeding probiotics, including following components: lactobacillus acidophilus, cud Bifidobacterium, candida utili, zytase, glucose oxidase, bacillus amyloliquefaciens, bacillus licheniformis, bacillus subtilis, lipase, acidic cellulase, acid protease.The bacillus amyloliquefaciens hypopus microbial dry powder: spore content >=1011A/gram；The bacillus licheniformis hypopus microbial dry powder: spore content >=1011A/gram；The bacillus subtilis hypopus microbial dry powder: spore content >=1011A/gram.The present invention significantly improves the eggshell quality of egg, reduces the yield of broken soft-shelled egg；Improve the egg production of the red laying hen in capital.It is obviously improved the formed stools of laying hen, hence it is evident that mitigate the ammonia taste of henhouse.
Description
Technical field
The invention belongs to fowl raising probiotics more particularly to a kind of layer breeding probiotics and its preparations
Method.
Background technique
In recent years, China's livestock and poultry breeding industry rapidly develop, scale intensive culture other than bringing high yield high benefit,
Also negative effect is brought, the use of a large amount of feed addictives, antibiotic, hormone medicine leads to medicament residue, causes livestock and poultry
Contamination of products.
Long-term a large amount of abuse of antibiotics accelerates the generation of a large amount of antibody-resistant bacterium in livestock and poultry cultivation, and animal body is caused to be exempted from
The decline of epidemic disease power induces the generation and prevalence of various epidemic diseases.Antibiotic preventive and therapeutic effect weakens significantly or even inefficacy, causes antibiosis
Element seriously remains in animal products, enters human body by food chain, seriously endangers the health of human body, and pollution of ecological environment is broken
The bad ecological balance threatens sanitarian safety.
Relatively common Substitutes For Antibiotic has probiotics, plant extracts, Chinese herb feed additive on the market at present
Equal products.Wherein probiotics to have no toxic side effect, have no drug resistance, noresidue, low cost, significant effect the features such as, gradually
The approval of numerous culturists and Consumption of animal by-product person are obtained, the developing direction of novel fodder additive is represented.
A kind of preparation method of young age animal and fowl fodder of CN201110421495.1, using enzymatic hydrolysis and fermentation asynchronous technique production
High nutrition, the prebiotic feed of young age livestock and poultry glycopeptide easy to digest by corn, wheat (or wheat-middlings, flour), are cracked rice, soybean and dregs of beans etc.
Five kinds of feedstuff impurity eliminations crush, addition water is modulated into enzymatic hydrolysis slurries after mixing, and a- amylase, saccharification is successively added
Enzyme, zytase, dextranase and microbial protease, component A is made in drying after enzymatic hydrolysis.Water is added in dregs of beans impurity elimination, is puddled
Inoculation yeast, lactic acid bacteria and bacillus licheniformis after uniformly, while microbial protease is added, it is dried after anaerobic fermentation and enzymatic hydrolysis
B component is made.Feed is made after two kinds of components are mixed in proportion.The feed is rich in short chain dextrin, glucose, oligosaccharide, small
Peptide, lactic acid and probiotics, have that amino acid balance, that nutrition is high, easy to digest, palatability is good and anti-nutritional factors content is low etc. is special
Point.
A kind of preparation method of additive for farm animal feed of CN201510658005.8 comprising following steps: S1, strain sieve
Choosing: in a variety of strains screening can be improved immunity of livestock probiotics strain it is stand-by；S2, single culture renewal cultivation, work
Change, expanding propagation；S3, multi-cultur es composite fermentation；S4, carrier is added to composite enzyme obtained in S3 and viable bacteria；S5, drying,
It crushes, obtains finished product, and test to finished product.Additive for farm animal feed provided by the invention is trained using the intersection of microorganism
The technology of supporting, by unique mixed fermentation technology continuous three times, to excite synzyme, catabolic enzyme, vitamin, the antibiosis of microorganism
Substance, growth hormone etc. are made feed addictive using the secretion and viable bacteria of these microorganisms, belong to pure biological agent, safety
Environmental protection, can be improved the immunity of livestock and poultry, and reduce feed utilization rate, control feed cost.
CNCN201210361014.7 one kind is solvable, young age digestible animal feed preparation method, corn, wheat and broken
Rice impurity elimination crushing and water-adding is modulated into slurries, and enzymolysis liquid is made by the way that a- amylase and carbohydrase is added.By filtrate and blood plasma egg
The mixing such as white, lactose are sized mixing, and dextrin concentrated sugar liquid is made.Filter residue, bean cake powder and water be inoculated with after mixing mushroom anaerobic fermentation and
Enzymatic hydrolysis, pulverizes, and small peptide protein milk is made in defibrination.It is mixed and made into pancebrin, cream is made after filters pressing, high pressure homogenizer homogeneous
Change liquid.Emulsion, which is directly packed, is made paste or liquid-state feed, or is made via popcorn absorption and tube bundle drying machine drying
Mash feed, or add water it is quenched after, by spray dryer drying little particle feed is made.Feed rich in short chain dextrin, glucose,
Oligosaccharide, small peptide, lactic acid and probiotics, with amino acid balance, nutrition is high, easy to digest, palatability is good and anti-nutritional factors contains
Measure low advantage.
There is also following technological deficiencies for the probiotics of the above-mentioned prior art: (1) do not improve significantly egg eggshell quality
Effect, broken soft-shelled egg has generation more, influences the quality of egg.(2) improvement is laid eggs not to the red laying hen in the capital of primiparity egg
Obviously, because it is digested and assimilated, function is not perfect, and many nutrition are unable to good absorption.(3) laying hen cannot preferably be improved
Digestive system flora, layer manure is thin shapeless, and ammonia taste is serious in henhouse.
Summary of the invention
In view of the above technical problems, the present invention provides a kind of layer breeding probiotics and preparation method thereof, to reach
To following goal of the invention: (1) improving the eggshell quality of egg, reduce the yield of broken soft-shelled egg；(2) production of the red laying hen in capital is improved
Egg amount.(3) formed stools of laying hen are obviously improved, hence it is evident that mitigate the ammonia taste of henhouse.
To reach the above goal of the invention, the technical scheme is that
A kind of layer breeding probiotics, in parts by weight, raw material includes:
10-20 parts of bacillus amyloliquefaciens hypopus microbial dry powder
5-15 parts of bacillus licheniformis hypopus microbial dry powder
10-20 parts of bacillus subtilis hypopus microbial dry powder
10-20 parts of lactobacillus acidophilus hypopus microbial dry powder
2-8 parts of hypopus microbial dry powder of cud Bifidobacterium
5-20 parts of candida utili hypopus microbial dry powder
1-10 parts of lipase
1-5 parts of zytase
1-5 parts of acidic cellulase
2-8 parts of glucose oxidase
3-8 parts of acid protease；
The deposit number of the bacillus amyloliquefaciens is ACCC10270；The deposit number of the bacillus licheniformis is
CICC20514；The deposit number of bacillus subtilis is CICC20517.
A kind of preparation method of layer breeding probiotics:
(1) bacillus amyloliquefaciens hypopus microbial dry powder is prepared
The preparation of the bacillus amyloliquefaciens ACCC10270 hypopus microbial dry powder the following steps are included:
Bacillus amyloliquefaciens strain on the slant medium of purifying culture is seeded to the work for being 15~30% equipped with volume ratio
In the conical flask for changing culture medium, the formula of the activation medium is beef-protein medium, and cultivation temperature is 35~38
DEG C, shaking flask 140~180rpm of revolving speed cultivates 10~12h；
The good bacillus amyloliquefaciens strain of activation culture is inoculated into the seed equipped with volume ratio for 50~70% fermentation mediums
In tank, volume inoculum concentration is 0.5~2%, and it is that 50~70% fermentations are trained that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
In the fermentor for supporting base, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is
20~35g/L of corn flour, 15~25g/L of dregs of beans, 0.5~2g/L of potassium dihydrogen phosphate, 1~3g/L of sodium chloride, dipotassium hydrogen phosphate 0.1
~1g/L, 0.5~1.5g/L of magnesium sulfate are 35~38 DEG C in cultivation temperature, 200~220rpm of fermentor speed of agitator culture 20
~32h, bacillus amyloliquefaciens gemma conversion ratio is 95% or more in fermentor, content >=1010A/ml, then obtains fermentation
Bacillus amyloliquefaciens it is spray-dried be prepared into hypopus microbial dry powder, bacillus amyloliquefaciens spore content in dry powder
≥1011A/gram.
(2) bacillus licheniformis hypopus microbial dry powder is prepared
The preparation of the bacillus licheniformis CICC20514 hypopus microbial dry powder the following steps are included:
Bacillus licheniformis strain on the slant medium of purifying culture is seeded to the activation for being 15~30% equipped with volume ratio
In the conical flask of culture medium, the formula of the activation medium is beef-protein medium, and cultivation temperature is 36~40 DEG C,
140~160rpm of shaking flask revolving speed cultivates 8~12h；
The good Bacillus licheniformis strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 50~70% fermentation mediums
Interior, volume inoculum concentration is 0.5~3%, and it is 50~70% fermented and cultureds that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
In the fermentor of base, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is jade
15~25g/L of rice flour, 2~10g/L of glucose, 15~25g/L of dregs of beans, 0.1~1g/L of dipotassium hydrogen phosphate, potassium dihydrogen phosphate 0.5
0.01~0.1g/L of~1.5g/L, 0.05~0.2g/L of calcium chloride and manganese sulfate is 36~40 DEG C in cultivation temperature, and fermentor stirs
It mixes 190~220rpm of revolving speed and cultivates 24~38h, bacillus licheniformis gemma conversion ratio is 95% or more in fermentor, content >=8
×109Then a/ml is prepared into hypopus microbial dry powder, dry powder for the obtained bacillus licheniformis of fermentation is spray-dried
Middle bacillus licheniformis spore content >=1011A/gram.
(3) the hypopus microbial dry powder of bacillus subtilis is prepared
The preparation of the bacillus subtilis CICC20517 hypopus microbial dry powder the following steps are included:
Bacillus subtilis strain on the slant medium of purifying culture is seeded to the activation for being 15~30% equipped with volume ratio
In the shaking flask of culture medium, the formula of the activation medium be 8~15g/L of peptone, 3~7g/L of sodium chloride and powdered beef 3~
8g/L is 36~40 DEG C in cultivation temperature, and 140~180rpm of shaking flask revolving speed cultivates 8~12h；
The good Bacillus subtilis strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 50~70% fermentation mediums
Interior, volume inoculum concentration is 0.2~1%, and it is 50~70% fermented and cultureds that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
In the fermentor of base, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is jade
20~30g/L of rice flour, 2~10g/L of glucose, 15~25g/L of beancake powder, 0.5~2g/L of ammonium sulfate, 1~5g/ of dipotassium hydrogen phosphate
L, 0.1~0.5g/L of 0.5~2g/L of magnesium sulfate and manganese sulfate is 36~40 DEG C in cultivation temperature, fermentor speed of agitator 200~
220rpm cultivates 20~30h, and bacillus subtilis spore conversion ratio is 95% or more in fermentor, content >=1010A/ml, then
Hypopus microbial dry powder, bacillus subtilis in dry powder are prepared by the obtained bacillus subtilis single bacterium of fermentation is spray-dried
Bacterium spore content >=1011A/gram.
(4) lactobacillus acidophilus hypopus microbial dry powder is prepared
The preparation of the lactobacillus acidophilus hypopus microbial dry powder the following steps are included:
Lactobacillus acidophilus species on the slant medium of purifying culture are seeded to the activation for being 50~70% equipped with volume ratio to train
In the shaking flask for supporting base, the formula of the activation medium is MRS culture medium, is 35~38 DEG C in cultivation temperature, stationary culture 20
~28h；
The good Lactobacillus acidophilus species of activation culture are inoculated into the seeding tank equipped with volume ratio for 50~70% fermentation mediums
Interior, volume inoculum concentration is 1~3%, and it is 50~70% fermentation mediums that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
Fermentor in, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is grape
10~20g/L of sugar, 10~20g/L of peptone, 2~10g/L of powdered beef, 1~5g/L of diammonium hydrogen citrate, 1~6g/L of sodium acetate
It is 35~38 DEG C, fermentor 20~30h of stationary culture in cultivation temperature with 0.1~1g/L of magnesium sulfate, acidophilus cream bar in fermentor
Bacterial content >=109A/ml, protective agent is freeze-dried is prepared into suspend mode for the lactobacillus acidophilus single bacterium addition for then obtaining fermentation
Body microbial dry powder, lactobacillus acidophilus content >=10 in dry powder10A/gram.
(5) cud Bifidobacterium hypopus microbial dry powder is prepared
The preparation of the cud Bifidobacterium hypopus microbial dry powder the following steps are included:
Cud bifidobacterium species on the slant medium of purifying culture are seeded to the activation for being 50~70% equipped with volume ratio
In the anaerobism bottle of culture medium, the formula of the activation medium is 2~10g/L of glucose, 10~25g/L of casein peptone, yeast
1~10g/L of powder, 0.5~5g/L of dipotassium hydrogen phosphate, 0.1~1g/L of cysteine, 0.1~0.5g/L of calcium chloride, zinc sulfate 0.1
0.1~0.8g/L of~1g/L and magnesium chloride is 36~39 DEG C in cultivation temperature, 40~48h of stationary culture；
The good cud bifidobacterium species of activation culture are inoculated into the seeding tank equipped with volume ratio for 60~80% fermentation mediums
Interior, volume inoculum concentration is 1~3%, and it is 60~80% fermentation mediums that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
Fermentor in, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is grape
10~20g/L of sugar, 15~30g/L of casein peptone, 5~15g/L of yeast powder, 0.5~5g/L of dipotassium hydrogen phosphate, cysteine 0.1
~1g/L, 0.1~0.5g/L of calcium chloride, 0.1~0.8g/L of 0.5~3g/L of Tween 80,0.1~1g/L of zinc sulfate and magnesium chloride,
It is 36~39 DEG C, fermentor 44~52h of stationary culture in cultivation temperature, cud Bifidobacterium content >=10 in fermentor8A/
Ml, then will cud Bifidobacterium single bacterium addition protective agent that fermentation obtains be freeze-dried is prepared into hypopus microorganism and does
Powder, cud Bifidobacterium content >=10 in dry powder9A/gram.
(6) candida utili hypopus microbial dry powder is prepared
The preparation of the candida utili hypopus microbial dry powder the following steps are included:
Candida utili strain on the slant medium of purifying culture is seeded to the activation for being 15~30% equipped with volume ratio
In the conical flask of culture medium, the formula of the activation medium is YPD culture medium, and cultivation temperature is 28~30 DEG C, shaking flask revolving speed
120~150rpm cultivates 22~26h；
The good candida utili strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 50~70% fermentation mediums
Interior, volume inoculum concentration is 1~3%, and it is 50~70% fermentation mediums that cultured seeding tank strain, which is inoculated into equipped with volume ratio,
Fermentor in, volume inoculum concentration is 5~10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is pancreas egg
White 20~30g/L of peptone, 15~25g/L of glucose, 1~10g/L of corn pulp, 1~5g/L of potassium dihydrogen phosphate, dipotassium hydrogen phosphate 2~
10~20g/L of 8g/L and urea is 28~30 DEG C in cultivation temperature, 180~200rpm of fermentor speed of agitator culture 36~
44h, candida utili content >=2 × 10 in fermentor9A/ml, the candida utili for then obtaining fermentation is through fluidizing
Bed fluidized drying is prepared into hypopus microbial dry powder, candida utili content >=10 in dry powder10A/gram.
(7) probiotics are made
According to 10-20 parts of bacillus amyloliquefaciens hypopus microbial dry powder, bacillus licheniformis hypopus microbial dry powder 5-
15 parts, 10-20 parts of bacillus subtilis hypopus microbial dry powder, 10-20 parts of lactobacillus acidophilus hypopus microbial dry powder, tumor
2-8 parts of hypopus microbial dry powder of stomach Bifidobacterium, 5-20 parts of candida utili hypopus microbial dry powder, lipase 1-10
Part, 1-5 parts of zytase, 1-5 parts of acidic cellulase, 2-8 parts of glucose oxidase, the weight ratio that 3-8 parts of acid protease,
It is mixed to get layer breeding probiotics.
By adopting the above-described technical solution, the beneficial effects of the present invention are:
(1) probiotics of the present invention can be such that the eggshell quality of egg is obviously improved, hence it is evident that reduce soft egg and break egg quantity.112 days
At initial stage Egg Production of Laying Hens stage in age, due to the digestion and absorption function of itself is not perfect, eggshell quality is poor, it may occur that soft
Egg breaks egg situation.After 12-14 days of the present invention edible, the broken soft-shelled egg quantity in egg is zero.
(2) probiotics of the present invention to improve laying hen egg yield effect clearly.The red laying hen in 112 Tianjing of age in days
1150, after eating 16-18 days of the present invention, per day egg production improves 14 jin.
(3) it is found in breeding process, microorganism formulation of the present invention significantly improves the formed stools situation of laying hen, chicken manure table
It now dries, reduces the ammonia concentration of henhouse, the ammonia of henhouse is substantially reduced.
Specific embodiment
Below with reference to specific embodiment, the present invention is further explained.
A kind of layer breeding probiotics of embodiment 1
A kind of layer breeding probiotics, in parts by weight, raw material includes:
10 parts of bacillus amyloliquefaciens hypopus microbial dry powder
5 parts of bacillus licheniformis hypopus microbial dry powder
10 parts of bacillus subtilis hypopus microbial dry powder
15 parts of lactobacillus acidophilus hypopus microbial dry powder
5 parts of microbial dry powder of cud Bifidobacterium hypopus
5 parts of candida utili hypopus microbial dry powder
2 parts of lipase
3 parts of zytase
3 parts of acidic cellulase
2 parts of glucose oxidase
8 parts of acid protease；
The deposit number of the bacillus amyloliquefaciens is ACCC10270；The deposit number of the bacillus licheniformis is
CICC20514；The deposit number of bacillus subtilis is CICC20517.
A kind of preparation method of layer breeding probiotics:
(1) bacillus amyloliquefaciens hypopus microbial dry powder is prepared
The preparation of the bacillus amyloliquefaciens ACCC10270 hypopus microbial dry powder the following steps are included:
Bacillus amyloliquefaciens strain on the slant medium of purifying culture is seeded to the activation for being 15% equipped with volume ratio to train
In the conical flask for supporting base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 38 DEG C, and shaking flask turns
Fast 140rpm cultivates 12h；
The good bacillus amyloliquefaciens strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 50% fermentation medium
Interior, volume inoculum concentration is 0.5%, and cultured seeding tank strain is inoculated into the fermentation equipped with volume ratio for 70% fermentation medium
In tank, volume inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, the formula of culture medium be corn flour 20g/L,
Dregs of beans 25g/L, potassium dihydrogen phosphate 2g/L, sodium chloride 1g/L, dipotassium hydrogen phosphate 0.1g/L, magnesium sulfate 1.5g/L are in cultivation temperature
35 DEG C, fermentor speed of agitator 220rpm cultivates 20h, and bacillus amyloliquefaciens gemma conversion ratio is 95% or more in fermentor, contains
Amount >=1010Then a/ml is prepared into hypopus microbial dry powder for the obtained bacillus amyloliquefaciens of fermentation are spray-dried,
Bacillus amyloliquefaciens spore content >=10 in dry powder11A/gram.
(2) bacillus licheniformis hypopus microbial dry powder is prepared
The preparation of the bacillus licheniformis CICC20514 hypopus microbial dry powder the following steps are included:
Bacillus licheniformis strain on the slant medium of purifying culture is seeded to the activation culture for being 30% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 36 DEG C, shaking flask revolving speed
140rpm cultivates 12h；
By the good Bacillus licheniformis strain of activation culture be inoculated into equipped with volume ratio be 50% fermentation medium seeding tank in,
Volume inoculum concentration is 0.5%, and cultured seeding tank strain is inoculated into the fermentor equipped with volume ratio for 70% fermentation medium
Interior, volume inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 15g/L, grape
Sugared 2g/L, dregs of beans 15g/L, dipotassium hydrogen phosphate 0.1g/L, potassium dihydrogen phosphate 1.5g/L, calcium chloride 0.2g/L and manganese sulfate 0.01g/
L is 36 DEG C in cultivation temperature, and fermentor speed of agitator 190rpm is cultivated for 24 hours, bacillus licheniformis gemma conversion ratio in fermentor
It is 95% or more, content >=8 × 109Then a/ml is prepared into suspend mode for the obtained bacillus licheniformis of fermentation is spray-dried
Body microbial dry powder, bacillus licheniformis spore content >=10 in dry powder11A/gram.
(3) the hypopus microbial dry powder of bacillus subtilis is prepared
The preparation of the bacillus subtilis CICC20517 hypopus microbial dry powder the following steps are included:
Bacillus subtilis strain on the slant medium of purifying culture is seeded to the activation culture for being 15% equipped with volume ratio
In the shaking flask of base, the formula of the activation medium is peptone 8g/L, sodium chloride 3g/L and powdered beef 5g/L, in cultivation temperature
It is 36 DEG C, shaking flask revolving speed 180rpm cultivates 8h；
By the good Bacillus subtilis strain of activation culture be inoculated into equipped with volume ratio be 50% fermentation medium seeding tank in,
Volume inoculum concentration is 0.2%, and cultured seeding tank strain is inoculated into the fermentor equipped with volume ratio for 50% fermentation medium
Interior, volume inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 30g/L, grape
Sugared 10g/L, beancake powder 15g/L, ammonium sulfate 0.5g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate 0.5g/L and manganese sulfate 0.5g/L,
Cultivation temperature is 36 DEG C, and fermentor speed of agitator 220rpm cultivates 20h, and bacillus subtilis spore conversion ratio is in fermentor
95% or more, content >=1010Then a/ml is prepared into hypopus for the obtained bacillus subtilis single bacterium of fermentation is spray-dried
Microbial dry powder, bacillus subtilis spore content >=10 in dry powder11A/gram.
(4) lactobacillus acidophilus hypopus microbial dry powder is prepared
The preparation of the lactobacillus acidophilus hypopus microbial dry powder the following steps are included:
Lactobacillus acidophilus species on the slant medium of purifying culture are seeded to the activation medium for being 50% equipped with volume ratio
Shaking flask in, it is 35 DEG C in cultivation temperature that the formula of the activation medium, which is MRS culture medium, stationary culture 28h；
It is body in the seeding tank of 50% fermentation medium equipped with volume ratio that the good Lactobacillus acidophilus species of activation culture, which are inoculated into,
Product inoculum concentration is 3%, and it is body in the fermentor of 50% fermentation medium equipped with volume ratio that cultured seeding tank strain, which is inoculated into,
Product inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 10g/L, peptone
10g/L, powdered beef 10g/L, diammonium hydrogen citrate 5g/L, sodium acetate 6g/L and magnesium sulfate 1g/L are 35 DEG C in cultivation temperature, hair
Fermentation tank stationary culture 20h, lactobacillus acidophilus content >=10 in fermentor9A/ml, the lactobacillus acidophilus for then obtaining fermentation are single
Protective agent is freeze-dried is prepared into hypopus microbial dry powder for bacterium addition, lactobacillus acidophilus content >=10 in dry powder10A/gram.
(5) cud Bifidobacterium hypopus microbial dry powder is prepared
The preparation of the cud Bifidobacterium hypopus microbial dry powder the following steps are included:
Cud bifidobacterium species on the slant medium of purifying culture are seeded to the activation culture for being 70% equipped with volume ratio
In the anaerobism bottle of base, the formula of the activation medium is glucose 2g/L, casein peptone 25g/L, yeast powder 1g/L, phosphoric acid hydrogen
Dipotassium 0.5g/L, cysteine 0.1g/L, calcium chloride 0.5g/L, zinc sulfate 0.1g/L and magnesium chloride 0.1g/L, in cultivation temperature
It is 36 DEG C, stationary culture 48h；
By the good cud bifidobacterium species of activation culture be inoculated into equipped with volume ratio be 60% fermentation medium seeding tank in,
Volume inoculum concentration be 1%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 60% fermentation medium fermentor in,
Volume inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 20g/L, casein
Peptone 15g/L, yeast powder 5g/L, dipotassium hydrogen phosphate 5g/L, cysteine 0.1g/L, calcium chloride 0.1g/L, Tween 80 0.5g/L,
Zinc sulfate 0.1g/L and magnesium chloride 0.8g/L is 36 DEG C, fermentor stationary culture 44h in cultivation temperature, and cud is double in fermentor
Discrimination bacillus content >=108A/ml, the cud Bifidobacterium single bacterium for then obtaining fermentation add the freeze-dried preparation of protective agent
At hypopus microbial dry powder, cud Bifidobacterium content >=10 in dry powder9A/gram.
(6) candida utili hypopus microbial dry powder is prepared
The preparation of the candida utili hypopus microbial dry powder the following steps are included:
Candida utili strain on the slant medium of purifying culture is seeded to the activation culture for being 30% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is YPD culture medium, and cultivation temperature is 30 DEG C, shaking flask revolving speed 120rpm training
Support 22h；
By the good candida utili strain of activation culture be inoculated into equipped with volume ratio be 50% fermentation medium seeding tank in,
Volume inoculum concentration be 1%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 50% fermentation medium fermentor in,
Volume inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is tryptone 20g/L, grape
Sugared 15g/L, corn pulp 1g/L, potassium dihydrogen phosphate 1g/L, dipotassium hydrogen phosphate 2g/L and urea 10g/L are 28 DEG C in cultivation temperature,
Fermentor speed of agitator 180rpm cultivates 36h, candida utili content >=2 × 10 in fermentor9A/ml, then will fermentation
Obtained candida utili is prepared into hypopus microbial dry powder through fluidized bed fluidized drying, and candida utili contains in dry powder
Amount >=1010A/gram.
(7) probiotics are made
According to 10 parts of bacillus amyloliquefaciens hypopus microbial dry powder, 5 parts of bacillus licheniformis hypopus microbial dry powder is withered
Careless 10 parts of bacillus hypopus microbial dry powder, 15 parts of lactobacillus acidophilus hypopus microbial dry powder, cud Bifidobacterium stops
5 parts of dormancy body microbial dry powder, 5 parts of candida utili hypopus microbial dry powder are 2 parts of lipase, 3 parts of zytase, acid
3 parts of cellulase, 2 parts of glucose oxidase, the weight ratio that 8 parts of acid protease is mixed to get layer breeding Tiny ecosystem system
Agent.
A kind of layer breeding probiotics of embodiment 2
A kind of layer breeding probiotics, in parts by weight, raw material includes:
15 parts of bacillus amyloliquefaciens hypopus microbial dry powder
10 parts of bacillus licheniformis hypopus microbial dry powder
15 parts of bacillus subtilis hypopus microbial dry powder
16 parts of lactobacillus acidophilus hypopus microbial dry powder
2 parts of microbial dry powder of cud Bifidobacterium hypopus
12 parts of candida utili hypopus microbial dry powder
6 parts of lipase
2 parts of zytase
1 part of acidic cellulase
5 parts of glucose oxidase
5 parts of acid protease；
The deposit number of the bacillus amyloliquefaciens is ACCC10270；The deposit number of the bacillus licheniformis is
CICC20514；The deposit number of bacillus subtilis is CICC20517.
A kind of preparation method of layer breeding probiotics:
(1) bacillus amyloliquefaciens hypopus microbial dry powder is prepared
The preparation of the bacillus amyloliquefaciens ACCC10270 hypopus microbial dry powder the following steps are included:
Bacillus amyloliquefaciens strain on the slant medium of purifying culture is seeded to the activation for being 20% equipped with volume ratio to train
In the conical flask for supporting base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 36 DEG C, and shaking flask turns
Fast 160rpm cultivates 11h；
The good bacillus amyloliquefaciens strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 60% fermentation medium
Interior, volume inoculum concentration is 1.2%, and cultured seeding tank strain is inoculated into the fermentation equipped with volume ratio for 60% fermentation medium
In tank, volume inoculum concentration is 8%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 30g/L, beans
Dregs of rice 19g/L, potassium dihydrogen phosphate 1.5g/L, sodium chloride 2.2g/L, dipotassium hydrogen phosphate 0.6g/L, magnesium sulfate 1.1g/L are in cultivation temperature
It is 37 DEG C, fermentor speed of agitator 210rpm cultivates 27h, and bacillus amyloliquefaciens gemma conversion ratio is 95% or more in fermentor,
Content >=1010Then a/ml is prepared into the obtained bacillus amyloliquefaciens of fermentation are spray-dried hypopus microorganism and does
Powder, bacillus amyloliquefaciens spore content >=10 in dry powder11A/gram.
(2) bacillus licheniformis hypopus microbial dry powder is prepared
The preparation of the bacillus licheniformis CICC20514 hypopus microbial dry powder the following steps are included:
Bacillus licheniformis strain on the slant medium of purifying culture is seeded to the activation culture for being 22% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 38 DEG C, shaking flask revolving speed
150rpm cultivates 10h；
By the good Bacillus licheniformis strain of activation culture be inoculated into equipped with volume ratio be 60% fermentation medium seeding tank in,
Volume inoculum concentration be 2%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 60% fermentation medium fermentor in,
Volume inoculum concentration is 8%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 19g/L, glucose
7g/L, dregs of beans 20g/L, dipotassium hydrogen phosphate 0.7g/L, potassium dihydrogen phosphate 1.2g/L, calcium chloride 1.1g/L and manganese sulfate 0.6g/L,
It is 38 DEG C in cultivation temperature, fermentor speed of agitator 200rpm cultivates 30h, and bacillus licheniformis gemma conversion ratio is in fermentor
95% or more, content >=8 × 109Then a/ml is prepared into hypopus for the obtained bacillus licheniformis of fermentation is spray-dried
Microbial dry powder, bacillus licheniformis spore content >=10 in dry powder11A/gram.
(3) the hypopus microbial dry powder of bacillus subtilis is prepared
The preparation of the bacillus subtilis CICC20517 hypopus microbial dry powder the following steps are included:
Bacillus subtilis strain on the slant medium of purifying culture is seeded to the activation culture for being 15% equipped with volume ratio
In the shaking flask of base, the formula of the activation medium is peptone 11g/L, sodium chloride 5g/L and powdered beef 5g/L, in culture temperature
Degree is 38 DEG C, and shaking flask revolving speed 160rpm cultivates 10h；
By the good Bacillus subtilis strain of activation culture be inoculated into equipped with volume ratio be 60% fermentation medium seeding tank in,
Volume inoculum concentration is 0.7%, and cultured seeding tank strain is inoculated into the fermentor equipped with volume ratio for 60% fermentation medium
Interior, volume inoculum concentration is 8%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 26g/L, grape
Sugared 7g/L, beancake powder 19g/L, ammonium sulfate 1.3g/L, dipotassium hydrogen phosphate 3g/L, magnesium sulfate 1.5g/L and manganese sulfate 0.3g/L,
Cultivation temperature is 38 DEG C, and fermentor speed of agitator 210rpm cultivates 25h, and bacillus subtilis spore conversion ratio is in fermentor
95% or more, content >=1010Then a/ml is prepared into hypopus for the obtained bacillus subtilis single bacterium of fermentation is spray-dried
Microbial dry powder, bacillus subtilis spore content >=10 in dry powder11A/gram.
(4) lactobacillus acidophilus hypopus microbial dry powder is prepared
The preparation of the lactobacillus acidophilus hypopus microbial dry powder the following steps are included:
Lactobacillus acidophilus species on the slant medium of purifying culture are seeded to the activation medium for being 60% equipped with volume ratio
Shaking flask in, the formula of the activation medium is MRS culture medium, is 36 DEG C in cultivation temperature, stationary culture is for 24 hours；
It is body in the seeding tank of 60% fermentation medium equipped with volume ratio that the good Lactobacillus acidophilus species of activation culture, which are inoculated into,
Product inoculum concentration is 2%, and it is body in the fermentor of 60% fermentation medium equipped with volume ratio that cultured seeding tank strain, which is inoculated into,
Product inoculum concentration is 7%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 15g/L, peptone
16g/L, powdered beef 7g/L, diammonium hydrogen citrate 3g/L, sodium acetate 4g/L and magnesium sulfate 0.6g/L are 36.5 in cultivation temperature
DEG C, fermentor stationary culture 26h, lactobacillus acidophilus content >=10 in fermentor9A/ml, the acidophilus cream for then obtaining fermentation
Protective agent is freeze-dried is prepared into hypopus microbial dry powder for the addition of bacillus single bacterium, lactobacillus acidophilus content >=10 in dry powder10
A/gram.
(5) cud Bifidobacterium hypopus microbial dry powder is prepared
The preparation of the cud Bifidobacterium hypopus microbial dry powder the following steps are included:
Cud bifidobacterium species on the slant medium of purifying culture are seeded to the activation culture for being 60% equipped with volume ratio
In the anaerobism bottle of base, the formula of the activation medium is glucose 6g/L, casein peptone 18g/L, yeast powder 6g/L, phosphoric acid hydrogen
Dipotassium 3.5g/L, cysteine 0.6g/L, calcium chloride 0.3g/L, zinc sulfate 0.7g/L and magnesium chloride 0.5g/L, in cultivation temperature
It is 38 DEG C, stationary culture 45h；
By the good cud bifidobacterium species of activation culture be inoculated into equipped with volume ratio be 70% fermentation medium seeding tank in,
Volume inoculum concentration be 2%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 70% fermentation medium fermentor in,
Volume inoculum concentration is 8%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 16g/L, casein
Peptone 22g/L, yeast powder 10g/L, dipotassium hydrogen phosphate 3.5g/L, cysteine 0.5g/L, calcium chloride 0.3g/L, Tween 80 1.8g/
L, zinc sulfate 0.7g/L and magnesium chloride 0.5g/L is 37 DEG C, fermentor stationary culture 48h in cultivation temperature, cud in fermentor
Bifidobacterium content >=108A/ml, the cud Bifidobacterium single bacterium for then obtaining fermentation add the freeze-dried system of protective agent
It is standby at hypopus microbial dry powder, cud Bifidobacterium content >=10 in dry powder9A/gram.
(6) candida utili hypopus microbial dry powder is prepared
The preparation of the candida utili hypopus microbial dry powder the following steps are included:
Candida utili strain on the slant medium of purifying culture is seeded to the activation culture for being 22% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is YPD culture medium, and cultivation temperature is 29 DEG C, shaking flask revolving speed 140rpm training
It supports for 24 hours；
By the good candida utili strain of activation culture be inoculated into equipped with volume ratio be 60% fermentation medium seeding tank in,
Volume inoculum concentration be 2%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 60% fermentation medium fermentor in,
Volume inoculum concentration is 7%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is tryptone 26g/L, grape
Sugared 19g/L, corn pulp 7g/L, potassium dihydrogen phosphate 3g/L, dipotassium hydrogen phosphate 5g/L and urea 16g/L are 29 DEG C in cultivation temperature,
Fermentor speed of agitator 190rpm cultivates 40h, candida utili content >=2 × 10 in fermentor9A/ml, then will fermentation
Obtained candida utili is prepared into hypopus microbial dry powder through fluidized bed fluidized drying, and candida utili contains in dry powder
Amount >=1010A/gram.
(7) probiotics are made
According to 15 parts of bacillus amyloliquefaciens hypopus microbial dry powder, 10 parts of bacillus licheniformis hypopus microbial dry powder,
15 parts of bacillus subtilis hypopus microbial dry powder, 16 parts of lactobacillus acidophilus hypopus microbial dry powder, cud Bifidobacterium
2 parts of hypopus microbial dry powder, 12 parts of candida utili hypopus microbial dry powder, 6 parts of lipase, 2 parts of zytase, acid
1 part of cellulase of property, 5 parts of glucose oxidase, the weight ratio that 5 parts of acid protease is mixed to get layer breeding Tiny ecosystem
Preparation.
A kind of layer breeding probiotics of embodiment 3
A kind of layer breeding probiotics, in parts by weight, raw material includes:
20 parts of bacillus amyloliquefaciens hypopus microbial dry powder
15 parts of bacillus licheniformis hypopus microbial dry powder
20 parts of bacillus subtilis hypopus microbial dry powder
20 parts of lactobacillus acidophilus hypopus microbial dry powder
8 parts of microbial dry powder of cud Bifidobacterium hypopus
20 parts of candida utili hypopus microbial dry powder
10 parts of lipase
5 parts of zytase
5 parts of acidic cellulase
8 parts of glucose oxidase
3 parts of acid protease；
The deposit number of the bacillus amyloliquefaciens is ACCC10270；The deposit number of the bacillus licheniformis is
CICC20514；The deposit number of bacillus subtilis is CICC20517.
A kind of preparation method of layer breeding probiotics:
(1) bacillus amyloliquefaciens hypopus microbial dry powder is prepared
The preparation of the bacillus amyloliquefaciens ACCC10270 hypopus microbial dry powder the following steps are included:
Bacillus amyloliquefaciens strain on the slant medium of purifying culture is seeded to the activation for being 30% equipped with volume ratio to train
In the conical flask for supporting base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 35 DEG C, and shaking flask turns
Fast 180rpm cultivates 10h；
The good bacillus amyloliquefaciens strain of activation culture is inoculated into the seeding tank equipped with volume ratio for 70% fermentation medium
Interior, volume inoculum concentration is 2%, and cultured seeding tank strain is inoculated into the fermentor equipped with volume ratio for 50% fermentation medium
Interior, volume inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 35g/L, dregs of beans
15g/L, potassium dihydrogen phosphate 0.5g/L, sodium chloride 3g/L, dipotassium hydrogen phosphate 1g/L, magnesium sulfate 0.5g/L are 38 in cultivation temperature
DEG C, fermentor speed of agitator 200rpm cultivates 32h, and bacillus amyloliquefaciens gemma conversion ratio is 95% or more in fermentor, content
≥1010Then a/ml is prepared into hypopus microbial dry powder for the obtained bacillus amyloliquefaciens of fermentation are spray-dried, does
Bacillus amyloliquefaciens spore content >=10 in powder11A/gram.
(2) bacillus licheniformis hypopus microbial dry powder is prepared
The preparation of the bacillus licheniformis CICC20514 hypopus microbial dry powder the following steps are included:
Bacillus licheniformis strain on the slant medium of purifying culture is seeded to the activation culture for being 15% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is beef-protein medium, and cultivation temperature is 40 DEG C, shaking flask revolving speed
160rpm cultivates 8h；
By the good Bacillus licheniformis strain of activation culture be inoculated into equipped with volume ratio be 70% fermentation medium seeding tank in,
Volume inoculum concentration be 3%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 50% fermentation medium fermentor in,
Volume inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 25g/L, glucose
10g/L, dregs of beans 25g/L, dipotassium hydrogen phosphate 1g/L, potassium dihydrogen phosphate 0.5g/L, calcium chloride 0.05g/L and manganese sulfate 0.01g/L,
It is 40 DEG C in cultivation temperature, fermentor speed of agitator 220rpm cultivates 38h, and bacillus licheniformis gemma conversion ratio is in fermentor
95% or more, content >=8 × 109Then a/ml is prepared into hypopus for the obtained bacillus licheniformis of fermentation is spray-dried
Microbial dry powder, bacillus licheniformis spore content >=10 in dry powder11A/gram.
(3) the hypopus microbial dry powder of bacillus subtilis is prepared
The preparation of the bacillus subtilis CICC20517 hypopus microbial dry powder the following steps are included:
Bacillus subtilis strain on the slant medium of purifying culture is seeded to the activation culture for being 30% equipped with volume ratio
In the shaking flask of base, the formula of the activation medium is peptone 15g/L, sodium chloride 7g/L and powdered beef 8g/L, in culture temperature
Degree is 40 DEG C, and shaking flask revolving speed 140rpm cultivates 12h；
By the good Bacillus subtilis strain of activation culture be inoculated into equipped with volume ratio be 70% fermentation medium seeding tank in,
Volume inoculum concentration be 1%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 70% fermentation medium fermentor in,
Volume inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is corn flour 20g/L, glucose
2g/L, beancake powder 25g/L, ammonium sulfate 2g/L, dipotassium hydrogen phosphate 5g/L, magnesium sulfate 2g/L and manganese sulfate 0.1g/L, in culture temperature
Degree is 40 DEG C, and fermentor speed of agitator 200rpm cultivates 30h, and bacillus subtilis spore conversion ratio is 95% or more in fermentor,
Content >=1010Then a/ml is prepared into the obtained bacillus subtilis single bacterium of fermentation is spray-dried hypopus microorganism and does
Powder, bacillus subtilis spore content >=10 in dry powder11A/gram.
(4) lactobacillus acidophilus hypopus microbial dry powder is prepared
The preparation of the lactobacillus acidophilus hypopus microbial dry powder the following steps are included:
Lactobacillus acidophilus species on the slant medium of purifying culture are seeded to the activation medium for being 70% equipped with volume ratio
Shaking flask in, it is 38 DEG C in cultivation temperature that the formula of the activation medium, which is MRS culture medium, stationary culture 20h；
It is body in the seeding tank of 70% fermentation medium equipped with volume ratio that the good Lactobacillus acidophilus species of activation culture, which are inoculated into,
Product inoculum concentration is 1%, and it is body in the fermentor of 70% fermentation medium equipped with volume ratio that cultured seeding tank strain, which is inoculated into,
Product inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 20g/L, peptone
20g/L, powdered beef 2g/L, diammonium hydrogen citrate 1g/L, sodium acetate 1g/L and magnesium sulfate 0.1g/L are 38 DEG C in cultivation temperature,
Fermentor stationary culture 30h, lactobacillus acidophilus content >=10 in fermentor9A/ml, the lactobacillus acidophilus for then obtaining fermentation
Protective agent is freeze-dried is prepared into hypopus microbial dry powder for single bacterium addition, lactobacillus acidophilus content >=10 in dry powder10A/
Gram.
(5) cud Bifidobacterium hypopus microbial dry powder is prepared
The preparation of the cud Bifidobacterium hypopus microbial dry powder the following steps are included:
Cud bifidobacterium species on the slant medium of purifying culture are seeded to the activation culture for being 50% equipped with volume ratio
In the anaerobism bottle of base, the formula of the activation medium is glucose 10g/L, casein peptone 10g/L, yeast powder 10g/L, phosphoric acid
Hydrogen dipotassium 5g/L, cysteine 1g/L, calcium chloride 0.1g/L, zinc sulfate 1g/L and magnesium chloride 0.8g/L are 39 in cultivation temperature
DEG C, stationary culture 40h；
By the good cud bifidobacterium species of activation culture be inoculated into equipped with volume ratio be 80% fermentation medium seeding tank in,
Volume inoculum concentration be 3%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 80% fermentation medium fermentor in,
Volume inoculum concentration is 10%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is glucose 10g/L, casein
Peptone 30g/L, yeast powder 15g/L, dipotassium hydrogen phosphate 0.5g/L, cysteine 1g/L, calcium chloride 0.5g/L, Tween 80 3g/L, sulphur
Sour zinc 1g/L and magnesium chloride 0.1g/L is 39 DEG C, fermentor stationary culture 52h in cultivation temperature, cud bifid bar in fermentor
Bacterial content >=108A/ml, protective agent is freeze-dried is prepared into not for the cud Bifidobacterium single bacterium addition for then obtaining fermentation
Dormancy body microbial dry powder, cud Bifidobacterium content >=10 in dry powder9A/gram.
(6) candida utili hypopus microbial dry powder is prepared
The preparation of the candida utili hypopus microbial dry powder the following steps are included:
Candida utili strain on the slant medium of purifying culture is seeded to the activation culture for being 15% equipped with volume ratio
In the conical flask of base, the formula of the activation medium is YPD culture medium, and cultivation temperature is 28 DEG C, shaking flask revolving speed 150rpm training
Support 26h；
By the good candida utili strain of activation culture be inoculated into equipped with volume ratio be 70% fermentation medium seeding tank in,
Volume inoculum concentration be 3%, by cultured seeding tank strain be inoculated into equipped with volume ratio be 70% fermentation medium fermentor in,
Volume inoculum concentration is 5%, and the seeding tank is identical with the formula of fermentor, and the formula of culture medium is tryptone 30g/L, grape
Sugared 25g/L, corn pulp 10g/L, potassium dihydrogen phosphate 5g/L, dipotassium hydrogen phosphate 8g/L and urea 20g/L are 30 in cultivation temperature
DEG C, fermentor speed of agitator 200rpm cultivates 44h, candida utili content >=2 × 10 in fermentor9A/ml, then will hair
The candida utili that ferment obtains is prepared into hypopus microbial dry powder, candida utili in dry powder through fluidized bed fluidized drying
Content >=1010A/gram.
(7) probiotics are made
According to 20 parts of bacillus amyloliquefaciens hypopus microbial dry powder, 15 parts of bacillus licheniformis hypopus microbial dry powder,
20 parts of bacillus subtilis hypopus microbial dry powder, 20 parts of lactobacillus acidophilus hypopus microbial dry powder, cud Bifidobacterium
8 parts of hypopus microbial dry powder, 20 parts of candida utili hypopus microbial dry powder, 10 parts of lipase, 5 parts of zytase,
5 parts of acidic cellulase, 8 parts of glucose oxidase, the weight ratio that 3 parts of acid protease is mixed to get layer breeding with micro- life
State preparation.
4 effect experiment of embodiment
To verify the influence for adding probiotics of the present invention in layer chicken feed to chicken group's production performance, carried out in Changyi City
20 days feeding experiments by a definite date.It is specific as follows:
Experimental design:
Test period: on June 17th, 2018 to July 7
Test place: Changyi City, Shandong Province wolf port chicken house
Laying hen: the red laying hen in 112 Tianjing of age in days, common chicken group is tested, two groups of number such as is randomly divided into, respectively experimental group and right
According to group, every group chicken 1150.
Feeding capacity: every 125 grams/day of chicken.
Experimental group and control group do not use any antibiotic during experiment.
Diet Formula:
The experimental results are shown inthe following table:
As it can be seen that egg production integrally reaches in 20 days lays eggs after adding probiotics of the present invention in 112 age in days egg feedstuffs
Peak constantly rises with using the time to extend egg production, tests 16-18 days, and the per day egg production of 1150 chickens is than control
Group improves 14 jin.This result illustrate probiotics of the present invention to improve laying hen egg yield effect clearly.
Meanwhile 112 initial stages in age in days Egg Production of Laying Hens stage, due to the digestion and absorption function of itself is not perfect, egg
Shell is second-rate, it may occur that soft egg breaks egg situation.It can be seen that from this experimental result after adding probiotics of the present invention, soft egg
Broken egg situation significantly reduces, it is seen that the eggshell quality of egg is obviously improved.
Meanwhile after addition probiotics of the present invention, effectively inhibit Escherichia coli, the growth of the harmful bacterias such as salmonella,
The growth for promoting beneficial bacterium, improves the health status of laying hen.Found in practical breeding process, hence it is evident that improve the excrement of laying hen at
Shape, chicken manure show drying, reduce the ammonia concentration of henhouse, the ammonia taste of henhouse is substantially reduced.
Economic efficiency contrast table:
The result shows that after addition probiotics of the present invention, hence it is evident that improve the eggshell quality of laying hen, break soft-shelled egg and reduce；It improves
The production performance of laying hen, obvious to 112 days laying hen effects, laying hen egg yield is obviously improved, and daily benefit increases by 54.2 yuan；
The ammonia concentration of henhouse is substantially reduced, and excrement molding improves.
This is the results showed that probiotics of the present invention have apparent inhibiting effect to laying hen rate of broken eggs, hence it is evident that reduces
Laying hen death and culling rate, peak of laying eggs are unified；Treasured of laying eggs can effectively inhibit Escherichia coli, and the growth of the harmful bacterias such as salmonella promotees
Into the growth of beneficial bacterium, improve the health status of laying hen；Contained a variety of microbes producing cellulases, can be effectively supplemented endogenous enzymes in alimentary canal
Deficiency, improve efficiency of feed utilization, improve egg production；Multivitamin is supplemented, laying hen immunity function is improved, is improved disease-resistant and anti-
Stress ability improves the unification of egg-laying peak, significantly improves eggshell quality, deepens eggshell color, reduces rate of broken eggs；Obvious drop
Low henhouse harmful gas concentration improves breeding environment, reduces the disease incidence of respiratory disease.
The above results prove that the daily benefit of experimental group increases by 54.2 yuan of benefit more, significant effect.
In addition to specified otherwise, percentage of the present invention is mass percent, and the ratio is mass ratio.
Finally, it should be noted that the foregoing is only a preferred embodiment of the present invention, it is not intended to restrict the invention,
It, for those skilled in the art, still can be with although describing the invention in detail with reference to the foregoing embodiments
It modifies the technical solutions described in the foregoing embodiments or equivalent replacement of some of the technical features.It is all
Within the spirit and principles in the present invention, any modification, equivalent replacement, improvement and so on should be included in guarantor of the invention
Within the scope of shield.
Claims (10)
1. a kind of layer breeding probiotics, it is characterised in that: including following components: lactobacillus acidophilus, cud bifid bar
Bacterium, candida utili, zytase, glucose oxidase.
2. a kind of layer breeding probiotics according to claim 1, it is characterised in that: including following components: solution
Bacillus amyloliquefaciens, bacillus licheniformis, bacillus subtilis, lipase, acidic cellulase, acid protease.
3. a kind of layer breeding probiotics according to claim 1, it is characterised in that: in parts by weight, including
Following components:
10-20 parts of bacillus amyloliquefaciens hypopus microbial dry powder
5-15 parts of bacillus licheniformis hypopus microbial dry powder
10-20 parts of bacillus subtilis hypopus microbial dry powder
10-20 parts of lactobacillus acidophilus hypopus microbial dry powder
2-8 parts of hypopus microbial dry powder of cud Bifidobacterium
5-20 parts of candida utili hypopus microbial dry powder
1-10 parts of lipase
1-5 parts of zytase
1-5 parts of acidic cellulase
2-8 parts of glucose oxidase
3-8 parts of acid protease.
4. a kind of layer breeding probiotics according to claim 1, it is characterised in that: the solution starch gemma bar
Bacterium hypopus microbial dry powder: spore content >=1011A/gram；The bacillus licheniformis hypopus microbial dry powder: gemma contains
Amount >=1011A/gram；The bacillus subtilis hypopus microbial dry powder: spore content >=1011A/gram；The acidophilus cream
Bacillus hypopus microbial dry powder: bacterial content >=1010A/gram；The cud Bifidobacterium hypopus microbial dry powder: bacterial content
≥109A/gram；The candida utili hypopus microbial dry powder: amount of yeast >=1010A/gram.
5. a kind of layer breeding probiotics according to claim 1, it is characterised in that: the solution starch gemma bar
The deposit number of bacterium is ACCC10270；The deposit number of the bacillus licheniformis is CICC20514；The preservation of bacillus subtilis
Number be CICC20517.
6. a kind of preparation method of layer breeding probiotics according to claim 1, it is characterised in that: including system
Standby lactobacillus acidophilus hypopus microbial dry powder: the formula of the fermentation medium used is 10~20g/L of glucose, peptone 10
~20g/L, 2~10g/L of powdered beef, 0.1~1g/L of 1~5g/L of diammonium hydrogen citrate, 1~6g/L of sodium acetate and magnesium sulfate, hair
Ferment cultivation temperature is 35~38 DEG C, and fermented incubation time is 20~30h, obtains lactobacillus acidophilus content >=10 in fermentor9A/
ml；The lactobacillus acidophilus single bacterium that fermentation obtains is prepared into hypopus microbial dry powder, lactobacillus acidophilus content >=10 in dry powder10
A/gram.
7. a kind of preparation method of layer breeding probiotics according to claim 1, it is characterised in that: further include
Preparation prepares cud Bifidobacterium hypopus microbial dry powder: including activation, seed culture, fermentation, freeze-drying step；It is described
Activation: the formula of the activation medium used for 2~10g/L of glucose, 10~25g/L of casein peptone, 1~10g/L of yeast powder,
0.5~5g/L of dipotassium hydrogen phosphate, 0.1~1g/L of cysteine, 0.1~0.5g/L of calcium chloride, 0.1~1g/L of zinc sulfate and chlorination
0.1~0.8g/L of magnesium；Activation culture temperature is 36~39 DEG C, and the activation culture time is 40~48h.
8. a kind of preparation method of layer breeding probiotics according to claim 6, it is characterised in that: described kind
Son culture: the good cud bifidobacterium species of activation culture are inoculated into the kind equipped with volume ratio for 60~80% fermentation mediums
In sub- tank, volume inoculum concentration is 1~3%；The fermentation: by cultured seeding tank strain be inoculated into equipped with volume ratio be 60~
In the fermentor of 80% fermentation medium, volume inoculum concentration is 5~10%；It is 36~39 DEG C in cultivation temperature, fermentor stands training
Support 44~52h, cud Bifidobacterium content >=10 in fermentor8A/ml, the cud Bifidobacterium single bacterium that fermentation is obtained are added
Protective agent is freeze-dried to be prepared into hypopus microbial dry powder, cud Bifidobacterium content >=10 in dry powder9A/gram.
9. a kind of preparation method of layer breeding probiotics according to claim 6, it is characterised in that: described kind
The culture medium prescription that son culture and fermentation step use is equal are as follows: 10~20g/L of glucose, 15~30g/L of casein peptone, yeast powder
5~15g/L, 0.5~5g/L of dipotassium hydrogen phosphate, 0.1~1g/L of cysteine, 0.1~0.5g/L of calcium chloride, Tween 80 0.5~
0.1~0.8g/L of 3g/L, 0.1~1g/L of zinc sulfate and magnesium chloride.
10. a kind of preparation method of layer breeding probiotics according to claim 1, it is characterised in that: also wrap
Include and prepare candida utili hypopus microbial dry powder: the formula of the fermentation medium used for 20~30g/L of tryptone,
15~25g/L of glucose, 1~10g/L of corn pulp, 1~5g/L of potassium dihydrogen phosphate, 2~8g/L of dipotassium hydrogen phosphate and urea 10~
20g/L, fermented and cultured temperature are 28~30 DEG C, fermentor 180~200rpm of speed of agitator, and fermented incubation time is 36~44h.
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| CN110731426A (en) * | 2019-11-27 | 2020-01-31 | 重庆工商大学 | fungal enzyme composition for feeding chicken and method for producing low-adhesive chicken manure |
| CN110973356A (en) * | 2019-12-28 | 2020-04-10 | 河南德邻生物制品有限公司 | Additive for feed capable of replacing antibiotics and using method thereof |
| CN111374237A (en) * | 2020-04-10 | 2020-07-07 | 聊城创新畜禽养殖有限公司 | Laying hen microecological compound preparation for egg producing period and preparation method and application thereof |
| CN111387366A (en) * | 2020-05-12 | 2020-07-10 | 博益德（北京）生物科技有限公司 | Laying hen heat stress resistant fermented feed and preparation method and application thereof |
| CN113812530A (en) * | 2021-09-18 | 2021-12-21 | 四川省畜牧科学研究院 | Multifunctional feed, preparation method and application thereof in meat rabbit stress resistance |
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| CN111374237A (en) * | 2020-04-10 | 2020-07-07 | 聊城创新畜禽养殖有限公司 | Laying hen microecological compound preparation for egg producing period and preparation method and application thereof |
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| CN113812530A (en) * | 2021-09-18 | 2021-12-21 | 四川省畜牧科学研究院 | Multifunctional feed, preparation method and application thereof in meat rabbit stress resistance |
| CN113812530B (en) * | 2021-09-18 | 2023-06-23 | 四川省畜牧科学研究院 | Multifunctional feed, preparation method and application thereof in resisting stress of meat rabbits |
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