CN102550815A - Fermented feed additive, preparation method and application - Google Patents

Fermented feed additive, preparation method and application Download PDF

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CN102550815A
CN102550815A CN2011104176235A CN201110417623A CN102550815A CN 102550815 A CN102550815 A CN 102550815A CN 2011104176235 A CN2011104176235 A CN 2011104176235A CN 201110417623 A CN201110417623 A CN 201110417623A CN 102550815 A CN102550815 A CN 102550815A
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mixed
bacterial classification
bacillus
medium
feed additive
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CN102550815B (en
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潘玉林
刘玉江
王永刚
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Shandong Zhongke Jiayi Bio Engineering Co ltd
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    • YGENERAL 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
    • Y02TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02PCLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
    • Y02P60/00Technologies relating to agriculture, livestock or agroalimentary industries
    • Y02P60/80Food processing, e.g. use of renewable energies or variable speed drives in handling, conveying or stacking
    • Y02P60/87Re-use of by-products of food processing for fodder production

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Abstract

The invention discloses a fermented feed additive, a preparation method and application thereof. The fermented feed additive is prepared by fermenting mixed strains and mixed auxiliary materials consisting of monosodium glutamate protein, bean pulp, bran, cottonseed meal, corn fibers, rice bran and powdered rice hulls, wherein the mixed strains are obtained by mixing activated strains of lactobacillus acidophilus, lactobacillus planetarium, lactobacillus casei, saccharomyces cerevisiae, bacillus subtilis and bacillus licheniformis in a proportion of (1-5): (1-5): (1-5): (1-5): (1-5): (1-5); and the dosage of the fermented feed additive is 5 to 100 percent of the weight of the animal feed when used for the animal feed. According to the fermented feed additive, the bean pulp, the bran, the cottonseed meal and other raw auxiliary materials are sufficiently utilized, so that the food consumption in the animal feeding process is reduced; the fermented feed additive can be used to partially or completely replace the animal feed, and antibiotic medicaments can be reduced or prevented from being used; and no medicament is left, so that the feeding cost of farmers is reduced.

Description

A kind of feed additive, preparation method and application
Technical field
The present invention relates to technical field of animal, relate in particular to a kind of feed additive.
Background technology
Improving constantly of Along with people's living standard, people are increasingly high to the attention of food quality and food security.A series of incidents such as the melamine that particularly took place in recent years, clenbuterol hydrochloride cause the attention of people to food security all the more.Therefore, the quality as the animal feed in the source of animal food seems particularly important.At present, in the animal feeding process, need constantly to add girth growth additives such as antibiotic, hormone, not only mouthfeel is very poor to cause the meat of producing, and contains a lot of medicament residues, and the edible back of people increases people's the resistance to the action of a drug, influences the healthy of people.Therefore,, utilize the bioengineering means to produce a kind of antibiotic fermented feed technology that do not contain, become the inexorable trend of feedstuff industry for the quality of production and the minimizing medicament residue that strengthens aquaculture.But be limited to the restriction of fermented bacterium and zymotechnique, the fermented feed technology is difficult to healthy and rapid development.
The high speed development of feedstuff industry makes feedstuff industry increasingly high to the demand of grain raw material, even the people occurs and livestock is fought for phenomenons such as the grain energy.In order to relax the situation that people and animals disaccord, made full use of the secondary material of raw material, reduce feed cost and aquaculture cost, utilize secondary material development fermented feed to become a kind of trend.
Summary of the invention
First technical problem to be solved by this invention is: to the deficiency that prior art exists, provide a kind of can reduce antibiotic use with stress, enhance immune power, reduce aquaculture cost and improve the feed additive of animal meat taste.
Second technical problem to be solved by this invention is: to the deficiency of prior art existence; Provide a kind of can reduce antibiotic use with stress, enhance immune power, production cost is low, reduce aquaculture cost and improve the preparation method of the feed additive of animal meat taste.
The 3rd technical problem to be solved by this invention is: provide a kind of can reduce antibiotic use with stress, enhance immune power, the application of feed additive in animal feed that reduces aquaculture cost and improve the animal meat taste.
For solving above-mentioned first technical problem, technical scheme of the present invention is:
A kind of feed additive; Formulated with the mixed accessories fermentation that comprises following raw material by mixed bacteria: monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls, said mixed bacteria are that the bacterial classification of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis that activation culture is obtained obtains according to the mixed of 1~5:1~5:1~5:1~5:1~5:1~5.
Preferably, said mixed accessories comprises that following raw materials by weight percent is formulated:
Monosodium glutamate albumen 8~15%,
Dregs of beans 6~10%,
Wheat bran 10~20%,
Cotton dregs 8~12%,
Zein fiber 15~25%,
Rice bran 14~28%,
Powdered rice hulls 20~35%.
For solving above-mentioned second technical problem, technical scheme of the present invention is:
A kind of preparation method of feed additive may further comprise the steps:
(1) preparation of mixed bacteria: under aseptic condition; Respectively in bacterial classification inoculation to the lactic acid bacteria activation medium with said Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei; In bacterial classification inoculation to the saccharomycete activation medium with said S. cervisiae; In bacterial classification inoculation to the bacillus activation medium with said bacillus subtilis and bacillus licheniformis, 36~38 ℃ of activation culture 42~54 hours; In the bacterial classification inoculation of the Lactobacillus acidophilus who again activation culture is obtained, lactobacillus plantarum and Lactobacillus casei to the lactic acid bacteria liquid culture medium; In bacterial classification inoculation to the saccharomycete liquid culture medium with said S. cervisiae; In bacterial classification inoculation to the bacillus liquid culture medium with said bacillus subtilis and bacillus licheniformis, 36~38 ℃ of enlarged culture 68~82 hours; Then in bacterial classification inoculation to the lactic acid bacteria liquid culture medium with the first time Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei that enlarged culture obtains; In bacterial classification inoculation to the saccharomycete liquid culture medium with said S. cervisiae; In bacterial classification inoculation to the bacillus liquid culture medium with said bacillus subtilis and bacillus licheniformis; Inoculum concentration is a volume ratio 8~12%; 36~38 ℃ of enlarged culture 42~54 hours for the second time, the bacterial classification of said bacillus subtilis and bacillus licheniformis carried out in the constant temperature shaking table during enlarged culture in the first time, under ventilation condition, carried out during enlarged culture in the second time; Each bacterial classification that the second time, enlarged culture obtained is obtained mixed bacteria according to the mixed of 1~5:1~5:1~5:1~5:1~5:1~5.
(2) mixed culture fermentation: monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls mixed obtaining mixed accessories respectively according to 8~15%, 6~10%, 10~20%, 8~12%, 15~25%, 14~28% and 20~35% weight ratio; Said mixed bacteria is diluted with deionized water according to the weight ratio of 1:4~6, and adding and dilution weight ratio are that the sugar of 1~2:50 obtains mixed bacteria liquid; With said mixed bacteria liquid according to 25~35% inoculation ratio; Be inoculated in the said mixed accessories and mixing and stirring; 24~26 ℃ fermented in fermenting cellar 5~7 days; Obtain the feed additive product, the quantity of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis bacterial classification difference>=0.3 * 10 in the said feed additive 8Individual/gram, 0.4 * 10 8Individual/gram, 0.2 * 10 8Individual/gram, 0.6 * 10 8Individual/gram, 0.5 * 10 8Individual/gram and 0.3 * 10 8Individual/gram.
Wherein, the prescription of said lactic acid bacteria activation medium is: peptone 10~12g/l, meat extract 10~12g/l, yeast extract 5~8g/l, K 2HPO 42~4g/l, Triammonium citrate 2~4g/l, sodium acetate 5~8g/l, glucose 20~25g/l, Tween80 1~2mL/l, MgSO 47H 2O 0.58~1g/l, MnSO 44H 2O 0.25~0.5g/l, agar 20~25g/l and CaCO 310~12g/l; The prescription of said saccharomycete activation medium is: potato 200~220g/l, glucose 20~25g/l and agar 20~25g/l; The prescription of said bacillus activation medium is: yeast soaks powder 0.7~0.9g/l, peptone 1~1.2g/l, glucose 1~1.2g/l, MgSO 40.2~0.3g/l, K 2HPO 41~1.2g/l, ammonium sulfate 0.2~0.3g/l and agar 20~25g/l.
The prescription of said lactic acid bacteria liquid culture medium is: peptone 10~12g/l, meat extract 10~12g/l, yeast extract 5~8g/l, K 2HPO 42~4g/l, Triammonium citrate 2~4g/l, sodium acetate 5~8g/l, glucose 20~25g/l, Tween80 1~2mL/l, MgSO 47H 2O 0.58~1g/l, MnSO 44H 2O 0.25~0.5g/l and CaCO 310~12g/l; The prescription of said yeast fluid nutrient medium is: potato 200~220g/l and glucose 20~25g/l; The prescription of said bacillus liquid culture medium is: yeast soaks powder 0.7~0.9g/l, peptone 1~1.2g/l, glucose 1~1.2g/l, MgSO 40.2~0.3g/l, K 2HPO 41~1.2g/l and ammonium sulfate 0.2~0.3g/l.
Preferably, when said lactic acid bacteria activation medium or lactic acid bacteria liquid medium preparation, said each culture medium prescription is mixed the back pH value be adjusted to 6.2~6.4,121 ℃ of sterilizations 15~20 minutes; When said saccharomycete activation medium or saccharomycete liquid medium preparation, said potato is prepared into potato juice mixes with each culture medium prescription again; When said bacillus activation medium or bacillus liquid medium preparation, said each culture medium prescription is mixed the back pH value be adjusted to 7.2~7.4,121 ℃ of sterilizations 15~20 minutes.
As a kind of improvement, after said mixed bacteria liquid and said mixed accessories mixed, put into fermenting cellar after packing with the fermented feed bag and ferment.Said fermented feed bag is the packaging film with unidirectional degassing function, has another name called the one-way exhaust valve bag, use said fermented feed bag packaged fermentation feed after, both guaranteed the ventilation of sweat, also guaranteed the germ-free condition of yeasting.Said fermented feed bag is the commercially available prod, and for example Cangnan county person of outstanding talent reaches the fermented feed bag that technology of the package factory produces.
The application of a kind of animal feed additive in animal feed, the addition when said feed additive is used as animal feed is 5~100wt% of animal feed.
Owing to adopted technique scheme, the invention has the beneficial effects as follows:
1, feed additive of the present invention; The bacterial classification that adopts Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis is as mixed bacteria; The bacterial classification of choosing can adapt to the gastric environment of strong acid rapidly, and the percent of pass and the field planting rate that get into enteron aisle are high; Can produce a large amount of probios, digestive ferment, B family vitamin, lysozyme, catalase, metabolic factor and other metabolites behind the said fermented by mixed bacterium; Probio can breed rapidly after getting into enteron aisle; Stimulate near the immune cell propagation of enteron aisle, in enteron aisle, can effectively control endotoxin, shigella dysenteriae, typhoid bacillus, paratyphosum Bacterium, campylobacter, staphylococcus etc. are had antagonism; The immunocompetence of enhancing body reduces the use of antibiotic medicine in the animal feeding process; Digestive ferment can promote the digest and decompose of feed, particularly to coarse-fibred decomposition utilization, improves the trans-utilization rate of feed; The growth of assorted bacterium in material such as lysozyme, catalase degradable or the inhibition enteron aisle, conditioning gut flora balance; Feed after the fermentation has unique ferment fragrance flavor, and food calling is effective, and palatability is good, can promote the animal feed.And the present invention can not discharge waste liquid and refuse in preparation process, reduced pollution and environmental protection treatment expense to environment.
2, the present invention is inoculated into mixed bacteria in the raw material auxiliary material and obtains after the fermentation, makes full use of raw material auxiliary materials such as dregs of beans, wheat bran, cotton dregs, has reduced the grain consumption in the animal-breeding process; Alleviated the predicament that people and animals disaccord; The feed additive of producing can add in the animal feed in proportion, also can all replace animal feed to use, and not only can reduce or avoid the use of the antibiotics medicine; Reduced raiser's aquaculture cost; Alleviated the breed workload, and the animal product delicious meat of producing, and do not have medicament residue.
3, sweat of the present invention ferments after adopting fermented feed bag packing, simplified fermentation condition, has reduced in the sweat problem that influences fermented quality because of living contaminants, has reduced in the sweat because of the waste of mildew factor to raw material.
The specific embodiment
Below in conjunction with concrete embodiment, further set forth the present invention.
Embodiment 1
Under aseptic condition; Respectively the bacterial classification of said Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei is scraped with transfer needle and got a ring; Be seeded in the culture dish that fills the lactic acid bacteria activation medium, said lactic acid bacteria activation medium is with peptone 10g/l, meat extract 10g/l, yeast extract 5g/l, K 2HPO 42g/l, Triammonium citrate 2g/l, sodium acetate 5g/l, glucose 20g/l, Tween80 1mL/l, MgSO 47H 2O 0.58g/l, MnSO 44H 2O 0.25g/l, agar 20g/l and CaCO 310g/l mixes the back pH value and is adjusted to 6.2, obtains in 15 minutes 121 ℃ of sterilizations; The bacterial classification of said S. cervisiae scraped with transfer needle get a ring; Be seeded in the culture dish that fills the saccharomycete activation medium, said saccharomycete activation medium is potato 200g/l to be prepared into obtain after potato juice, glucose 20g/l and agar 20g/l mix; The bacterial classification of said bacillus subtilis and bacillus licheniformis scraped with transfer needle get a ring; Be seeded in the culture dish that fills the bacillus activation medium, said bacillus activation medium is that yeast is soaked powder 0.7g/l, peptone 1g/l, glucose 1g/l, MgSO 40.2g/l, K 2HPO 41g/l, ammonium sulfate 0.2g/l and agar 20g/l mix the back pH value and are adjusted to 7.2, obtain in 15 minutes 121 ℃ of sterilizations; Each bacterial classification was 36 ℃ of activation culture 42 hours; The bacterial classification of the Lactobacillus acidophilus who again activation culture is obtained, lactobacillus plantarum and Lactobacillus casei is scraped with the aseptic inoculation pin and is got a pin; Be seeded in the triangular flask that fills lactic acid bacteria liquid culture medium; And triangular flask placed the constant temperature shaking table; The bacterial classification of said S. cervisiae scraped with the aseptic inoculation pin get a pin, be seeded in the triangular flask that fills saccharomycete liquid culture medium, the bacterial classification of said bacillus subtilis and bacillus licheniformis is scraped with the aseptic inoculation pin got a pin; Be seeded in the triangular flask that fills bacillus liquid culture medium, each bacterial classification was 36 ℃ of enlarged culture 68 hours; The bacterial classification of inciting somebody to action the first time Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei that enlarged culture obtains then is with being seeded in the seeding tank that fills lactic acid bacteria liquid culture medium; With the bacterial classification inoculation of said S. cervisiae to the seeding tank that fills saccharomycete liquid culture medium; With the bacterial classification inoculation of said bacillus subtilis and bacillus licheniformis to the seeding tank that fills bacillus liquid culture medium; Each bacterial classification inoculation amount is volume ratio 8%; 36 ℃ of enlarged culture 42 hours for the second time, wherein the enlarged culture need second time of bacillus subtilis and bacillus licheniformis carry out under ventilation condition.Said lactic acid bacteria liquid culture medium is with peptone 10g/l, meat extract 10g/l, yeast extract 5g/l, K 2HPO 42g/l, Triammonium citrate 2g/l, sodium acetate 5g/l, glucose 20g/l, Tween80 1mL/l, MgSO 47H 2O 0.58g/l, MnSO 44H 2O 0.25g/l and CaCO 310g/l mixes the back pH value and is adjusted to 6.2, obtains in 15 minutes 121 ℃ of sterilizations; Said yeast fluid nutrient medium is potato 200g/l to be prepared into obtain after potato juice mixes with glucose 20g/l then; Said bacillus liquid culture medium is that yeast is soaked powder 0.7g/l, peptone 1g/l, glucose 1g/l, MgSO 40.2g/l, K 2HPO 41g/l and ammonium sulfate 0.2g/l mix the back pH value and are adjusted to 7.2, obtain in 15 minutes 121 ℃ of sterilizations.Each bacterial classification that the second time, enlarged culture obtained is obtained mixed bacteria according to the mixed of 1:1:1:1:1:1.
Auxiliary material monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls mixed according to 8%, 6%, 10%, 8%, 15%, 14% and 20% weight ratio respectively obtain mixed accessories; Said mixed bacteria is diluted with deionized water according to the weight ratio of 1:4, and adding and dilution weight ratio are that the sugar of 1:50 obtains mixed bacteria liquid; With said mixed bacteria liquid according to 25% inoculation ratio; Be inoculated in the said mixed accessories and mixing and stirring; Put into fermenting cellar after packing with the fermented feed bag; 24 ℃ fermented 5 days in fermenting cellar, obtained the feed additive product, the quantity of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis bacterial classification difference>=0.3 * 10 in the said feed additive 8Individual/gram, 0.4 * 10 8Individual/gram, 0.2 * 10 8Individual/gram, 0.6 * 10 8Individual/gram, 0.5 * 10 8Individual/gram and 0.3 * 10 8Individual/gram.
Embodiment 2
Under aseptic condition; Respectively the bacterial classification of said Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei is scraped with transfer needle and got a ring; Be seeded in the culture dish that fills the lactic acid bacteria activation medium, said lactic acid bacteria activation medium is with peptone 12g/l, meat extract 12g/l, yeast extract 8g/l, K 2HPO 44g/l, Triammonium citrate 4g/l, sodium acetate 8g/l, glucose 25g/l, Tween80 2mL/l, MgSO 47H 2O 1g/l, MnSO 44H 2O 0.5g/l, agar 25g/l and CaCO 312g/l mixes the back pH value and is adjusted to 6.4, obtains in 20 minutes 121 ℃ of sterilizations; The bacterial classification of said S. cervisiae scraped with transfer needle get a ring; Be seeded in the culture dish that fills the saccharomycete activation medium, said saccharomycete activation medium is potato 220g/l to be prepared into obtain after potato juice, glucose 25g/l and agar 25g/l mix; The bacterial classification of said bacillus subtilis and bacillus licheniformis scraped with transfer needle get a ring; Be seeded in the culture dish that fills the bacillus activation medium, said bacillus activation medium is that yeast is soaked powder 0.9g/l, peptone 1.2g/l, glucose 1.2g/l, MgSO 40.3g/l, K 2HPO 41.2g/l, ammonium sulfate 0.3g/l and agar 25g/l mix the back pH value and be adjusted to 7.4, obtained in 20 minutes 121 ℃ of sterilizations; Each bacterial classification was 38 ℃ of activation culture 54 hours; The bacterial classification of the Lactobacillus acidophilus who again activation culture is obtained, lactobacillus plantarum and Lactobacillus casei is scraped with the aseptic inoculation pin and is got a pin; Be seeded in the triangular flask that fills lactic acid bacteria liquid culture medium; And triangular flask placed the constant temperature shaking table; The bacterial classification of said S. cervisiae scraped with the aseptic inoculation pin get a pin, be seeded in the triangular flask that fills saccharomycete liquid culture medium, the bacterial classification of said bacillus subtilis and bacillus licheniformis is scraped with the aseptic inoculation pin got a pin; Be seeded in the triangular flask that fills bacillus liquid culture medium, each bacterial classification was 38 ℃ of enlarged culture 82 hours; The bacterial classification of inciting somebody to action the first time Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei that enlarged culture obtains then is with being seeded in the seeding tank that fills lactic acid bacteria liquid culture medium; With the bacterial classification inoculation of said S. cervisiae to the seeding tank that fills saccharomycete liquid culture medium; With the bacterial classification inoculation of said bacillus subtilis and bacillus licheniformis to the seeding tank that fills bacillus liquid culture medium; Each bacterial classification inoculation amount is volume ratio 12%; 38 ℃ of enlarged culture 54 hours for the second time, wherein the enlarged culture need second time of bacillus subtilis and bacillus licheniformis carry out under ventilation condition.Said lactic acid bacteria liquid culture medium is with peptone 12g/l, meat extract 12g/l, yeast extract 8g/l, K 2HPO 44g/l, Triammonium citrate 4g/l, sodium acetate 8g/l, glucose 25g/l, Tween80 2mL/l, MgSO 47H 2O 1g/l, MnSO 44H 2O 0.5g/l and CaCO 312g/l mixes the back pH value and is adjusted to 6.4, obtains in 20 minutes 121 ℃ of sterilizations; Said yeast fluid nutrient medium is potato 220g/l to be prepared into obtain after potato juice mixes with glucose 25g/l then; Said bacillus liquid culture medium is that yeast is soaked powder 0.9g/l, peptone 1.2g/l, glucose 1.2g/l, MgSO 40.3g/l, K 2HPO 4Be adjusted to 7.4 1.2g/l mix the back pH value, obtained in 20 minutes 121 ℃ of sterilizations with ammonium sulfate 0.3g/l.Each bacterial classification that the second time, enlarged culture obtained is obtained mixed bacteria according to the mixed of 5:5:5:5:5:5.
Auxiliary material monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls mixed according to 15%, 10%, 20%, 12%, 25%, 28% and 35% weight ratio respectively obtain mixed accessories; Said mixed bacteria is diluted with deionized water according to the weight ratio of 1:6, and adding and dilution weight ratio are that the sugar of 2:50 obtains mixed bacteria liquid; With said mixed bacteria liquid according to 35% inoculation ratio; Be inoculated in the said mixed accessories and mixing and stirring; Put into fermenting cellar after packing with the fermented feed bag; 26 ℃ fermented 7 days in fermenting cellar, obtained the feed additive product, the quantity of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis bacterial classification difference>=0.3 * 10 in the said feed additive 8Individual/gram, 0.4 * 10 8Individual/gram, 0.2 * 10 8Individual/gram, 0.6 * 10 8Individual/gram, 0.5 * 10 8Individual/gram and 0.3 * 10 8Individual/gram.
Embodiment 3
Under aseptic condition; Respectively the bacterial classification of said Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei is scraped with transfer needle and got a ring; Be seeded in the culture dish that fills the lactic acid bacteria activation medium, said lactic acid bacteria activation medium is with peptone 11g/l, meat extract 11g/l, yeast extract 6g/l, K 2HPO 43g/l, Triammonium citrate 3g/l, sodium acetate 6g/l, glucose 22g/l, Tween80 1.5mL/l, MgSO 47H 2O 0.7g/l, MnSO 44H 2O 0.3g/l, agar 22g/l and CaCO 311g/l mixes the back pH value and is adjusted to 6.3, obtains in 15 minutes 121 ℃ of sterilizations; The bacterial classification of said S. cervisiae scraped with transfer needle get a ring; Be seeded in the culture dish that fills the saccharomycete activation medium, said saccharomycete activation medium is potato 210g/l to be prepared into obtain after potato juice, glucose 22g/l and agar 22g/l mix; The bacterial classification of said bacillus subtilis and bacillus licheniformis scraped with transfer needle get a ring; Be seeded in the culture dish that fills the bacillus activation medium, said bacillus activation medium is that yeast is soaked powder 0.8g/l, peptone 1.1g/l, glucose 1.1g/l, MgSO 40.25g/l, K 2HPO 41.1g/l, ammonium sulfate 0.25g/l and agar 22g/l mix the back pH value and be adjusted to 7.3, obtained in 15 minutes 121 ℃ of sterilizations; Each bacterial classification was 37 ℃ of activation culture 48 hours; The bacterial classification of the Lactobacillus acidophilus who again activation culture is obtained, lactobacillus plantarum and Lactobacillus casei is scraped with the aseptic inoculation pin and is got a pin; Be seeded in the triangular flask that fills lactic acid bacteria liquid culture medium; And triangular flask placed the constant temperature shaking table; The bacterial classification of said S. cervisiae scraped with the aseptic inoculation pin get a pin, be seeded in the triangular flask that fills saccharomycete liquid culture medium, the bacterial classification of said bacillus subtilis and bacillus licheniformis is scraped with the aseptic inoculation pin got a pin; Be seeded in the triangular flask that fills bacillus liquid culture medium, each bacterial classification was 37 ℃ of enlarged culture 72 hours; The bacterial classification of inciting somebody to action the first time Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei that enlarged culture obtains then is with being seeded in the seeding tank that fills lactic acid bacteria liquid culture medium; With the bacterial classification inoculation of said S. cervisiae to the seeding tank that fills saccharomycete liquid culture medium; With the bacterial classification inoculation of said bacillus subtilis and bacillus licheniformis to the seeding tank that fills bacillus liquid culture medium; Each bacterial classification inoculation amount is volume ratio 10%; 37 ℃ of enlarged culture 48 hours for the second time, wherein the enlarged culture need second time of bacillus subtilis and bacillus licheniformis carry out under ventilation condition.Said lactic acid bacteria liquid culture medium is with peptone 11g/l, meat extract 11g/l, yeast extract 6g/l, K 2HPO 43g/l, Triammonium citrate 3g/l, sodium acetate 6g/l, glucose 22g/l, Tween80 1.5mL/l, MgSO 47H 2O 0.6g/l, MnSO 44H 2O 0.3g/l and CaCO 311g/l mixes the back pH value and is adjusted to 6.3, obtains in 15~20 minutes 121 ℃ of sterilizations; Said yeast fluid nutrient medium is potato 200~220g/l to be prepared into obtain after potato juice mixes with glucose 22g/l then; Said bacillus liquid culture medium is that yeast is soaked powder 0.8g/l, peptone 1.1g/l, glucose 1.1g/l, MgSO 40.25g/l, K 2HPO 4Be adjusted to 7.3 1.1g/l mix the back pH value, obtained in 15 minutes 121 ℃ of sterilizations with ammonium sulfate 0.25g/l.Each bacterial classification that the second time, enlarged culture obtained is obtained mixed bacteria according to the mixed of 2:2:3:2:3:1.
Auxiliary material monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls mixed according to 10%, 8%, 15%, 10%, 20%, 20% and 25% weight ratio respectively obtain mixed accessories; Said mixed bacteria is diluted with deionized water according to the weight ratio of 1:5, and adding and dilution weight ratio are that the sugar of 1:50 obtains mixed bacteria liquid; With said mixed bacteria liquid according to 30% inoculation ratio; Be inoculated in the said mixed accessories and mixing and stirring; Put into fermenting cellar after packing with the fermented feed bag; 25 ℃ fermented 6 days in fermenting cellar, obtained the feed additive product, the quantity of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis bacterial classification difference>=0.3 * 10 in the said feed additive 8Individual/gram, 0.4 * 10 8Individual/gram, 0.2 * 10 8Individual/gram, 0.6 * 10 8Individual/gram, 0.5 * 10 8Individual/gram and 0.3 * 10 8Individual/gram.
Embodiment 4
The feed additive that embodiment 1, embodiment 2, embodiment 3 are obtained adds in the chicken feed according to 5% weight addition, and chicken is fed.
Embodiment 5
The feed additive that embodiment 1, embodiment 2, embodiment 3 are obtained adds in the chicken feed according to 20% weight addition, and chicken is fed.
Embodiment 6
The feed additive that embodiment 1, embodiment 2, embodiment 3 obtain is fed to chicken as the chicken feed.
The comparative example 1
Compare test; Test is carried out on the laying hen in 23 ages in week; To test laying hen and be divided into three groups: control group and one group of experiment, two groups of experiments; Wherein control group is fed with common chicken feed, tests one group of feed additive that obtains with embodiment 1 and embodiment 2 respectively with two groups of experiments and adds in the chicken feed according to 5% weight addition and feed, and experimentation does not all use the antibiotics medicine; Test period is from totally 91 days November 25,2010 of 27 days to 2010 August in, and experimental result sees the following form.
Figure DEST_PATH_883465DEST_PATH_IMAGE001
Can find out from above comparative test result; Do not using equally under the condition of antibiotics medicine; The one group of death rate with the laying hen of two groups of experiments of experiment that has added animal feed additive of the present invention is significantly less than the death rate of control group; Prove absolutely that the animal feed additive that the present invention prepares can improve animal immunizing power, under the prerequisite that strengthens addition, can avoid the use of the antibiotics medicine fully.

Claims (8)

1. feed additive; It is characterized in that by mixed bacteria formulated with the mixed accessories fermentation that comprises following raw material: monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls, said mixed bacteria are that the bacterial classification of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis that activation culture is obtained obtains according to the mixed of 1~5:1~5:1~5:1~5:1~5:1~5.
2. feed additive as claimed in claim 1 is characterized in that said mixed accessories comprises that following raw materials by weight percent is formulated:
Monosodium glutamate albumen 8~15%,
Dregs of beans 6~10%,
Wheat bran 10~20%,
Cotton dregs 8~12%,
Zein fiber 15~25%,
Rice bran 14~28%,
Powdered rice hulls 20~35%.
3. according to claim 1 or claim 2 the preparation method of feed additive is characterized in that may further comprise the steps:
(1) preparation of mixed bacteria: under aseptic condition; Respectively in bacterial classification inoculation to the lactic acid bacteria activation medium with said Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei; In bacterial classification inoculation to the saccharomycete activation medium with said S. cervisiae; In bacterial classification inoculation to the bacillus activation medium with said bacillus subtilis and bacillus licheniformis, 36~38 ℃ of activation culture 42~54 hours; In the bacterial classification inoculation of the Lactobacillus acidophilus who again activation culture is obtained, lactobacillus plantarum and Lactobacillus casei to the lactic acid bacteria liquid culture medium; In bacterial classification inoculation to the saccharomycete liquid culture medium with said S. cervisiae; In bacterial classification inoculation to the bacillus liquid culture medium with said bacillus subtilis and bacillus licheniformis, 36~38 ℃ of enlarged culture 68~82 hours; Then in bacterial classification inoculation to the lactic acid bacteria liquid culture medium with the first time Lactobacillus acidophilus, lactobacillus plantarum and Lactobacillus casei that enlarged culture obtains; In bacterial classification inoculation to the saccharomycete liquid culture medium with said S. cervisiae; In bacterial classification inoculation to the bacillus liquid culture medium with said bacillus subtilis and bacillus licheniformis; Inoculum concentration is a volume ratio 8~12%; 36~38 ℃ of enlarged culture 42~54 hours for the second time, the bacterial classification of said bacillus subtilis and bacillus licheniformis carried out in the constant temperature shaking table during enlarged culture in the first time, under ventilation condition, carried out during enlarged culture in the second time; Each bacterial classification that the second time, enlarged culture obtained is obtained mixed bacteria according to the mixed of 1~5:1~5:1~5:1~5:1~5:1~5;
(2) mixed culture fermentation: monosodium glutamate albumen, dregs of beans, wheat bran, cotton dregs, zein fiber, rice bran and powdered rice hulls mixed obtaining mixed accessories respectively according to 8~15%, 6~10%, 10~20%, 8~12%, 15~25%, 14~28% and 20~35% weight ratio; Said mixed bacteria is diluted with deionized water according to the weight ratio of 1:4~6, and adding and dilution weight ratio are that the sugar of 1~2:50 obtains mixed bacteria liquid; With said mixed bacteria liquid according to 25~35% inoculation ratio; Be inoculated in the said mixed accessories and mixing and stirring; 24~26 ℃ fermented in fermenting cellar 5~7 days; Obtain the feed additive product, the quantity of Lactobacillus acidophilus, lactobacillus plantarum, Lactobacillus casei, S. cervisiae, bacillus subtilis and bacillus licheniformis bacterial classification difference>=0.3 * 10 in the said feed additive 8Individual/gram, 0.4 * 10 8Individual/gram, 0.2 * 10 8Individual/gram, 0.6 * 10 8Individual/gram, 0.5 * 10 8Individual/gram and 0.3 * 10 8Individual/gram.
4. the preparation method of feed additive as claimed in claim 3, it is characterized in that: the prescription of said lactic acid bacteria activation medium is: peptone 10~12g/l, meat extract 10~12g/l, yeast extract 5~8g/l, K 2HPO 42~4g/l, Triammonium citrate 2~4g/l, sodium acetate 5~8g/l, glucose 20~25g/l, Tween80 1~2mL/l, MgSO 47H 2O 0.58~1g/l, MnSO 44H 2O 0.25~0.5g/l, agar 20~25g/l and CaCO 310~12g/l; The prescription of said saccharomycete activation medium is: potato 200~220g/l, glucose 20~25g/l and agar 20~25g/l; The prescription of said bacillus activation medium is: yeast soaks powder 0.7~0.9g/l, peptone 1~1.2g/l, glucose 1~1.2g/l, MgSO 40.2~0.3g/l, K 2HPO 41~1.2g/l, ammonium sulfate 0.2~0.3g/l and agar 20~25g/l.
5. the preparation method of feed additive as claimed in claim 3, it is characterized in that: the prescription of said lactic acid bacteria liquid culture medium is: peptone 10~12g/l, meat extract 10~12g/l, yeast extract 5~8g/l, K 2HPO 42~4g/l, Triammonium citrate 2~4g/l, sodium acetate 5~8g/l, glucose 20~25g/l, Tween80 1~2mL/l, MgSO 47H 2O 0.58~1g/l, MnSO 44H 2O 0.25~0.5g/l and CaCO 310~12g/l; The prescription of said yeast fluid nutrient medium is: potato 200~220g/l and glucose 20~25g/l; The prescription of said bacillus liquid culture medium is: yeast soaks powder 0.7~0.9g/l, peptone 1~1.2g/l, glucose 1~1.2g/l, MgSO 40.2~0.3g/l, K 2HPO 41~1.2g/l and ammonium sulfate 0.2~0.3g/l.
6. like the preparation method of claim 4 or 5 described feed additives; It is characterized in that: when said lactic acid bacteria activation medium or lactic acid bacteria liquid medium preparation; Said each culture medium prescription is mixed the back pH value be adjusted to 6.2~6.4,121 ℃ of sterilizations 15~20 minutes; When said saccharomycete activation medium or saccharomycete liquid medium preparation, said potato is prepared into potato juice mixes with each culture medium prescription again; When said bacillus activation medium or bacillus liquid medium preparation, said each culture medium prescription is mixed the back pH value be adjusted to 7.2~7.4,121 ℃ of sterilizations 15~20 minutes.
7. the preparation method of feed additive as claimed in claim 3 is characterized in that: after said mixed bacteria liquid and said mixed accessories are mixed, put into fermenting cellar after packing with the fermented feed bag and ferment.
8. according to claim 1 or claim 2 the application of feed additive in animal feed is characterized in that: the addition of said feed additive during as animal feed is 5~100wt% of animal feed.
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