CN108450664A - A kind of high-efficiency feed additive product - Google Patents

A kind of high-efficiency feed additive product Download PDF

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Publication number
CN108450664A
CN108450664A CN201810174306.7A CN201810174306A CN108450664A CN 108450664 A CN108450664 A CN 108450664A CN 201810174306 A CN201810174306 A CN 201810174306A CN 108450664 A CN108450664 A CN 108450664A
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bacillus subtilis
culture
product
ganoderma lucidum
animal
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邵素英
苏俊
王玉
李政
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Abstract

The invention discloses a kind of high-efficiency feed additive product, composition is as follows:Bacterium powder 20%, Ganoderma Lucidum solid fermentation culture 20% is lyophilized in oligofructose 10%, soya-bean polypeptides 10%, bacillus subtilis;Active dry yeast 10%;Bacterium powder 10% is lyophilized in Lactobacillus rhamnosus, bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.Product of the present invention can effectively reduce usage amount of the antibiotics in raising animal, improve the safety of animal meat product, improve the food utilization efficiency of animal, enhance the immunity of animal, improve raise benefit.

Description

A kind of high-efficiency feed additive product
The application is the divisional application of following application:The applying date is on 06 24th, 2012, application No. is 201210213402.0 a kind of entitled high-efficiency feed additive product.
Technical field:
The invention belongs to field of fodder more particularly to feed addictives.
Background technology:
During modern livestock and poultry cultivation, in order to prevent and treat Animal diseases, excessive antibiotic product is often used to ensure Animal health, but the use of excessive antibiotic often results in the quality decline of animal meat product and its product and antibiotic resistance increases By force, human health is caused also to be seriously affected by antibiotic problem by the transmission of food chain.
Some natural materials from nature not only have good nutritive peculiarity, while also have antiviral, anti-inflammatory, It is anti-oxidant, adjust body's immunity the effects that, have broad application prospects.
Soya-bean polypeptides absorbability easy to digest promotes energetic supersession, to show anti-obesic action;Promote over fatigue to restore The effect of spirit, muscle power;Increase the endurance of muscle, myoglobins is promoted to restore;There is the effects that low antigen, hypoallergenic.
Ganoderma lucidum [Ganoderema lucidum (Leyss ex Fr.) karst], also known as polyporus lucidus, category Basidiomycotina, Aphyllophorales, Ganodermataceae, Ganoderma.Doctor's allusion quotation in successive dynasties playbacks ganoderma lucidum in " medicine-feeding ", ranks before ginseng.《The legendary god of farming Book on Chinese herbal medicine warp》、《Compendium of Materia Medica》It records, ganoderma lucidum materials for improving vision, tonifying liver gas, tranquilizing the mind, strengthening the essence, invigorating heart gas, beneficial temper, beneficial lung qi, kidney-nourishing Gas, logical nine orifices, ear is clever, mesh is bright, sharp joint, diuresis, skin maintenance, be first-class excellent tonic product.Ganoderma lucidum polysaccharide can promote albumen The synthesis of matter, nucleic acid has facilitation to update, the synthesis of serum, liver and bone marrow cell protein or nucleic acid, it is spirit One of the principle active component that sesame is strengthened the body resistance to consolidate the constitution.Ganoderma lucidum polysaccharide is primarily present in lucid ganoderma fungus fructification, sclerotium, mycelium or hair In zymotic fluid;The ganoderic acid being present in ganoderma lucidum has analgesic, calmness, inhibits histamine releasing, removing toxic substances, liver protection, poisoning tumour cell Function, be one of principle active component of ganoderma lucidum.
How effective exploitation obtains green feed additive product with natural plants or fungi is sent out feed additive industry Exhibition is of great significance.
Invention content:
The present invention provides a kind of feedstuff additive product;
Composition is as follows:
Bacterium powder 20%, ganoderma lucidum bacteria solid fermentation culture is lyophilized in oligofructose 10%, soya-bean polypeptides 20%, bacillus subtilis Bacterium powder 10%, bacillus subtilis (Bacillus is lyophilized in object 30%, active dry yeast 10%, Lactobacillus rhamnosus Subtilis it is) CGMCC6012, aforementioned proportion is mass percent.
Ganoderma Lucidum solid fermentation culture preparation method is as follows:
Slant strains activation culture:The ganoderma lucidum slant strains of preservation are transferred on slant medium, 22-28 DEG C of culture 96-120 hours or so, until mycelia covers with inclined-plane;25 DEG C of optimum culturing temperature.
1. liquid first order seed culture:By the access of one piece of above-mentioned ganoderma lucidum slant strains picking equipped with 100 milliliters of culture mediums First order seed culture, condition of culture are carried out in 500 milliliters of shaking flasks:80-180 revs/min of rotary shaker, 22-28 DEG C of culture 96- 130 hours or so;150 revs/min of rotary shaker is advisable, and cultivation temperature is advisable with 25 DEG C.
2. liquid two stage seed culture:Level-one shake-flask seed is equipped with 100 milliliters of culture mediums with the access of 5-20% inoculum concentrations 500 milliliters of shaking flasks in carry out secondary seed culture, condition of culture:80-180 revs/min of rotary shaker, 22-28 DEG C of culture 96-130 hours or so;Suitable inoculum concentration is 10%, and 150 revs/min of rotary shaker is advisable, and cultivation temperature is advisable with 25 DEG C.
3. solid fermentation culture:By second-level shake flask seed with the access of 5-10% inoculum concentrations equipped with solid fermentation culture after sterilizing In the 500L solid-state fermentation tanks of base, condition of culture:Charge 50%, 20-27 DEG C of cultivation temperature, humidity 75-90%, ventilation quantity 0.3-1.0 (V/V), incubation time 15-40 days.Stirring 10 minutes was opened every 3-10 days.Using SGF solid-state fermentation tanks.
4. fermented and cultured terminates, select a variety of drying means such as fluid bed, by dry materials to moisture 7% with Under.Solid fermentation material is crushed.
The carbon source of solid fermentation culture medium of the present invention mainly selects wheat bran, glucose, sucrose, starch, corn flour, corncob The common raw material such as (after crushing), nitrogen source can select groundnut meal, peptone, De-fatted soya protein Powder, beancake powder, yeast powder Deng;Inorganic salts to be added are needed to have potassium dihydrogen phosphate, magnesium sulfate.
After being crushed for 2-6% added with mass ratio in above-mentioned solid fermentation culture medium, Radix Angelicae Sinensis, the 3-8% of 40-50 mesh are crossed Crushing after, cross 40-50 mesh Radix Isatidis, 2-5% crush after, cross 40-50 mesh thyme.
The bacillus subtilis be bacillus subtilis CGMCC6012, be lyophilized bacterium powder in number of viable be (4-8) × 108A/g.The composite feed additive is made by following methods:
Bacterium powder etc. is lyophilized proportionally in Ganoderma Lucidum culture, oligofructose, soya-bean polypeptides and bacillus subtilis after crushing It is mixed to get feed addictive.
Soya-bean polypeptides use commercially available soya-bean polypeptides product, and molecular weight is between 500Da~700Da.
The present invention provides a kind of Feed Manufacturing bacillus subtilis (Bacillus subtilis) strain, and the bacterial strain is Through being preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address position on April 16th, 2012 In Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, (postcode 100101), deposit number is CGMCC No.6012.
Saccharomyces cerevisiae (Saccharomyces cerevisiae) provided by the present invention, the bacterial strain are preserved in the micro- life of China Object culture presevation administration committee common micro-organisms center, deposit number are CGMCC No.4429, preservation address:Court of Beijing The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica, postcode 100101.Preservation date December 08 in 2010 Day.
The bacterial strain feature is as follows:
It observes under the microscope, the cell of the bacterial strain is oval, one end budding, and size is about 1 × 5 μm;In solid culture On base, which is milky, and surface is smooth, and moistening, sticky, edge is more neat and medium bigger than normal.It, should compared with original bacteria Mutagenic strain is morphologically significantly less than starting strain.
Starting strain saccharomyces cerevisiae CICC31481 is purchased from Chinese industrial Microbiological Culture Collection administrative center.
Yeast strain of the present invention carries out selection and breeding using following flows:
The original strain that sets out → test tube activation → dithyl sulfate (DES) mutagenesis → hypertonic plate screening → nitrosoguanidine (NTG) mutagenesis screening → hypertonic tablet primary dcreening operation → shaking flask secondary screening → mitotic stability experiment → 7L fermentation tank tests
The present invention first uses DES to carry out mutagenesis to starting strain, passes through the hypertonic tablet (150g/L of brewer's wort after mutagenesis KCl) culture medium primary dcreening operation then continues NTG mutagenesis to the bacterial strain selected, passes through the hypertonic tablet (200g/L of brewer's wort KCl then) culture medium primary dcreening operation uses 250mL triangular flask secondary screenings, the excellent yeast strain of selection and breeding then to do passage assays, evaluate Its genetic stability is used in combination liquid chromatogram, gas chromatograph, gas chromatography mass spectrometry to measure the metabolite content in zymotic fluid, is finally adopted The evaluation of experiment effect is carried out with 7L fermentation tanks.
Bacterial strain CGMCC No.4429 genetic stabilities the result shows that:By continuous passage ten times, property indices all compare Relatively stablize, heredity is preferable, and character is not replied, therefore the purpose bacterial strain that bacterial strain CGMCC No.4429 are obtained as selection and breeding.
Purpose bacterial strain CGMCC No.4429 are done into the experiment of 7L fermentation tanks, the results showed that:Compared with starting strain, CGMCC No.4429 initial glucoses tolerable concentration can reach 300g/L, and 50% is improved compared with original bacteria;After fermentation, residual Sugar is 0.5g/L, glycerol content 1g/L, and 10% is improved compared with original bacteria;Acetic acid content is 0.8g/L, lactic acid content is 0.4g/L, basic and original bacteria are suitable;Concentration of alcohol is 175g/L, and 108% is improved compared with original bacteria.
Lactobacillus rhamnosus preparation process is:
Lactobacillus rhamnosus powdery bacterium powder production stage is as follows:Slant strains are transferred to fluid nutrient medium and are spread cultivation step by step To requirement
Volume;The bacterium solution to spread cultivation is centrifuged, precipitation thalline is collected;Protection is added into precipitation thalline Agent is simultaneously
It is diluted;Powdery microbial inoculum is prepared using drying equipment, Lactobacillus rhamnosus is (1-9) × 109A/gram
Lactobacillus rhamnosus used medium is as follows:Slant medium (g/L):20 glucose, 5 yeast powders, 10 soybean eggs White peptone, 10 beef extracts, 5NaCl, 10 sodium acetates, 2 ammonium citrates, 0.2MgSO4·7H2O, 0.05MnSO4·7H2O, 15 agar, pH 6.5。
Fermentation medium (g/L):40 glucose, 10 yeast powders, 10 soy peptones, inorganic salts (0.01NaCl, 0.5 second Sour sodium, 0.2 ammonium citrate, 0.2KH2PO4,0.2MgSO4·7H2O,0.05MnSO4·7H2O), 6.5 pH.
Active dry yeast is prepared using conventional production process, and active dry yeasr is (1-10) × 10 in active dry yeast powder9A/ Gram.
Advantageous effect:
The present invention is compounded by science, bacillus subtilis, Lactobacillus rhamnosus, yeast and Ganoderma Lucidum is realized organic Combination, is especially added to traditional Chinese medicine ingredients, product of the present invention can effectively reduce antibiotics and raise in Ganoderma Lucidum culture The usage amount in animal is supported, the safety of animal meat product is improved, improves the food utilization efficiency of animal, enhances the immune of animal Power improves raise benefit.
Specific implementation method:
Ganoderma Lucidum culture production process is as follows in the present invention:
(1) slant strains activation culture:
(2) liquid first order seed culture:
(3) liquid two stage seed culture:
(4) solid fermentation culture:
(5) fermented and cultured terminates, and selects a variety of drying means such as fluid bed, by dry materials to moisture 7% with Under.Solid fermentation material is crushed.
Lucidum strain used in the present invention is purchased from Chinese industrial Culture Collection, bacterium CICC14080.
Slant medium composition can be PDA slant mediums or other suitable culture mediums in the present invention.
The carbon source of solid fermentation culture medium of the present invention mainly selects wheat bran, glucose, sucrose, starch, corn flour, corncob The common raw material such as (after crushing), nitrogen source can select groundnut meal, peptone, De-fatted soya protein Powder, beancake powder, yeast powder Deng;Inorganic salts to be added are needed to have potassium dihydrogen phosphate, magnesium sulfate.
It is the crushing that 2-6% smashes it through the Radix Angelicae Sinensis of 40-50 mesh, 3-8% to be also added with mass ratio in the fermentation medium The Radix Isatidis of 40-50 mesh is crossed afterwards, and 2-5% smashes it through the thyme of 40-50 mesh;
Liquid fermentation seed culture medium forms in the present invention:Starch 2-3%, sucrose 3-5%, glucose 1-3%, peptone 0.5%, De-fatted soya protein Powder 2-3%, potassium dihydrogen phosphate 0.13-0.2%, magnesium sulfate 0.1-0.15%, vitamin B1 2PPm, PH6-7.
Ganoderma lucidum cultural method, which is suitble to belong to other strains and passes through fermentation method, in the present invention carries out large scale fermentation culture.
Induction mutation of bacterium
Take ultraviolet mutagenesis method:Using CICC10023 as starting strain, bacteria suspension is made, is coated on culture medium plate, It is cultivated in 37 DEG C of incubators after ultraviolet irradiation 90s.
Bacterial screening
30 single bacterium colonies for growing strong are chosen from 90s plates, are inoculated on inclined-plane, and in 37 DEG C of incubators, culture 24 is small When, 0--4 DEG C is stored refrigerated;As a contrast with original strain, using conventional herd breeding condition, sodium carboxymethylcellulose is as carbon source With proteolysis circle method screen cellulase-producing and the strong bacterial strain of protease, screening obtains cellulase-producing and protease is strong Strains B. subtilis (Bacillus subtilis) L1, this bacterial strain cellulase-producing ability are improved than original strain 25%, production protease ability improves 52% than original strain.The strain is entrusted in China General Microbiological culture presevation management Member's meeting common micro-organisms center preservation, deposit number are CGMCC No 6012.
Bacillus subtilis used in this product (Bacillus subtilis) strain, the bacterial strain is April 16 in 2012 It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center day, preservation address is located at Chaoyang District, Beijing City north The institute 3 of occasion West Road 1, (postcode 100101), deposit number are CGMCC No6012.
Lactobacillus rhamnosus (Lactobacillus rhamnosus) CGMCC No.4430 bacterial strain features are as follows:Micro- Under the microscope, which is rod-shaped, and width is less than 1 μm, and 2 to 3 bacillus, which are easy to be linked to be, to link together;In solid medium On, which is milky, and surface is smooth, and moistening, sticky, edge is more neat.Compared with original bacteria, the mutagenic strain is in shape It is significantly less than starting strain in state.Starting strain Lactobacillus rhamnosus CGMCC No.1.2134 are purchased from Chinese microorganism strain guarantor Hide administration committee's common micro-organisms center.Lactobacillus rhamnosus of the present invention carries out selection and breeding using following flows:It is original go out bacterium germination Sugared plate screening → nitrosoguanidine (NTG) mutagenesis of kind → test tube activation → high temperature acclimation → dithyl sulfate (DES) mutagenesis → height Screening → high temperature bacterium screening → shaking flask secondary screening → mitotic stability experiment → 5L fermentation tank tests.By purpose bacterial strain CGMCC No.4430 does the experiment of 5L lactic acid fermentation tanks, the results showed that:Compared with starting strain, CGMCC No.4430 glucose-tolerant concentration 270g/L can be reached, 95% is improved compared with original bacteria;After fermentation, lactic acid content 60g/L, compared with original bacteria Improve 158%.
Lactobacillus rhamnosus (Lactobacillus rhamnosus) CGMCC No.4430 depositary institutions are Chinese common Microbiological Culture Collection administrative center, the deposit date is on December 08th, 2010.Address:BeiChen West Road, Chaoyang District, BeiJing City 1 Institute 3, Institute of Microorganism, Academia Sinica, postcode 100101.
The performance of product of the present invention is stablized, safe to use, with other feed addictives without incompatibility.Product of the present invention Additive amount in feed is 1-2%, it completely can be with substitute antibiotics drug, and noresidue is free from environmental pollution.After feeding, The immunity and premunition of animal body can be significantly increased, growth is promoted, improves daily gain and feed conversion rate, and has anti-answer Swash, antioxidant effect;There is stronger prevention effect to intestinal bacteriosis simultaneously.According to experiment, feeding feed addition of the present invention The aquatic products of agent, quality are improved, and meet the greenization production requirement of animal food, Social benefit and economic benefit is very Significantly.
For the Chinese tradition edible mushroom that product of the present invention is produced using biological fermentation process as raw material, product quality is stably and controllable, production Product in vivo noresidue, non-environmental-pollution, without drug resistance, and immunity can be improved, with strong bacteriostasis, Neng Gouti High culture efficiency.Product complies fully with the requirement of green feed additive.
Since ganoderma lucidum has higher nutritive value and bacteria resistance function, the immunity of animal, and noresidue can be enhanced, It is environmentally safe.Ganoderma lucidum is produced by biofermentation technique and is processed as feed addictive, complies fully with current feed The developing direction of additive is a kind of green feed additive.
Embodiment 1 (feedstuff additive product)
It is as follows that the present invention provides a kind of feedstuff additive product composition:
Bacterium powder 20%, ganoderma lucidum bacteria solid fermentation culture is lyophilized in oligofructose 10%, soya-bean polypeptides 20%, bacillus subtilis Object 30%;Active dry yeast 10%;Bacterium powder 10%, bacillus subtilis (Bacillus is lyophilized in Lactobacillus rhamnosus Subtilis it is) CGMCC6012, aforementioned proportion is mass percent.Lactobacillus rhamnosus is CGMCC No.4430, and activity is dry Yeast is CGMCC No.4429.
Oligofructose is commercial product.
Ganoderma Lucidum solid fermentation culture preparation method is as follows:
Slant strains activation culture:The ganoderma lucidum slant strains of preservation are transferred on slant medium, 25 DEG C of cultures 110 are small When or so, until mycelia covers with inclined-plane;
1. liquid first order seed culture:By the access of one piece of above-mentioned ganoderma lucidum slant strains picking equipped with 100 milliliters of culture mediums First order seed culture, condition of culture are carried out in 500 milliliters of shaking flasks:100 revs/min of rotary shaker, 25 DEG C of cultures, 100 hours left sides It is right;25 DEG C of cultivation temperature.
2. liquid two stage seed culture:By level-one shake-flask seed with the access of 10% inoculum concentration equipped with 100 milliliters of culture mediums Secondary seed culture, condition of culture are carried out in 500 milliliters of shaking flasks:100 revs/min of rotary shaker, 25 DEG C of cultures, 130 hours left sides It is right;25 DEG C of cultivation temperature.
3. solid fermentation culture:By second-level shake flask seed with the access of 8% inoculum concentration equipped with solid fermentation culture medium after sterilizing 500L solid-state fermentation tanks in, condition of culture:Charge 50%, 25 DEG C of cultivation temperature, humidity 85%, ventilation quantity 0.5 (V/V), Incubation time 25 days.Stirring 10 minutes was opened every 4 days.Using SGF solid-state fermentation tanks.
4. fermented and cultured terminates, fluidized bed drying method is selected, by dry materials to moisture below 7%.To solid Body fermentation materials crush.
Solid fermentation culture medium composition is as follows in the present invention:60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corn Core (after crushing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% yeast powder;3% smash it through 50 mesh Radix Angelicae Sinensis, 4% Radix Isatidis for smashing it through 40 mesh, 3% smashes it through the thyme of 40 mesh;Medium pH is natural.Sterilising conditions:121 DEG C, 2 hours.
The bacillus subtilis is bacillus subtilis CGMCC6012, and it is 6 × 10 that number of viable in bacterium powder, which is lyophilized,8A/ g。
The composite feed additive is made by following methods:
Bacterium powder etc. is lyophilized proportionally in Ganoderma Lucidum culture, oligofructose, soya-bean polypeptides and bacillus subtilis after crushing It is mixed to get feed addictive.
Soya-bean polypeptides sell soya-bean polypeptides product using Heilungkiang flood icing the more health food Co., Ltd.
Product using effect
Using effect experiment of 1 feed addictive of example of the present invention in weanling pig
Test method:
Experimental animal selects Ningxia feed corporation,Ltd subordinate farm average weight for the strong of (7.16 ± 0.15) kg Health weanling pig 120, statistical analysis weight differences are not notable.Using single-factor Randomized Designs, 120 health are weaned Piglet, it is fifty-fifty by male and female, it is divided into 2 groups (control group and test groups), every group of 6 repetitions, each repeats 10 pigs.Test group is added Product of the present invention made from embodiment 1, control group do not add product of the present invention.Continuous feeding 30 days uses compared with the control group Invention feed addictive can get following effect:Test result shows to add this product in weanling pig daily ration, the day of piglet Weightening is improved a lot than control group;Feedstuff-meat ratio is substantially reduced;Diarrhea rate reduces 70.7%.
Using effect experiment of 1 feed addictive of example of the present invention in milking sow
21 days by a definite date feeding experiments have been carried out on Ningxia feed corporation,Ltd subordinate pig farm.Experiment uses single factor test pair Than design, 30 milking sows healthy, farrowing head number and birth counterpoise are similar, parity is 2 or 3 tires are randomly selected, It is randomly divided into 2 groups (i.e. test groups and control group), every group of 15 repetitions.Wherein:The addition of test group daily ration is prepared by embodiment 2 This product, control group do not add product of the present invention.Record weaned piglet weight, diarrhea rate, the death rate.Experiments have shown that:Test group 42.5% can be improved than control group weight of weaning litter, piglet head daily gain improves 31.6%, and diarrhea rate reduces 51.3%, the death rate Reduce by 60.3%.
The above results show that product of the present invention can improve sow and piglet body immunity, reduce antibiotic dosage, increase Add cultivation quality and benefits.

Claims (1)

1. a kind of high-efficiency feed additive product, composition is as follows:Oligofructose 10%, soya-bean polypeptides 20%, bacillus subtilis Bacterium powder 20%, Ganoderma Lucidum solid fermentation culture 30% is lyophilized;Active dry yeast 10%;Bacterium powder is lyophilized in Lactobacillus rhamnosus 10%;
The bacillus subtilis (Bacillus subtilis) is CGMCC6012;
The active dry yeast is CGMCC No.4429;
In the ganoderma lucidum bacteria solid fermentation used medium Radix Angelicae Sinensis of 40-50 mesh, 3- are smashed it through for 2-6% added with mass ratio 8% Radix Isatidis for smashing it through 40-50 mesh, 2-5% smash it through the thyme of 40-50 mesh;
The number of viable of the Lactobacillus rhamnosus freeze-drying bacterium powder is (1-9) × 109A/gram;The number of viable of active dry yeast reaches (1-10)×109A/gram;
Aforementioned proportion is mass percent.
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CN108220208B (en) * 2018-03-16 2020-06-12 青岛农业大学 Zinc-rich bacterial strain and application thereof
CN109362948A (en) * 2018-10-23 2019-02-22 南宁市黄陈生猪养殖场 A method of Se-enriched feedstuff additive is prepared using Moringa waste wood

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Application publication date: 20180828