CN102657287B - Compound feed addictive with bacteria and oxidation resisting functions - Google Patents
Compound feed addictive with bacteria and oxidation resisting functions Download PDFInfo
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Abstract
The invention relates to a compound feed addictive with bacteria and oxidation resisting functions. The compound feed addictive is prepared from the following components in percentage by weight: 10-15% of antibacterial peptide, 10-20% of propolis, 10-25% of fructus ligustri lucidi extractant, 15-30% of lyophilized bacillus subtilis bacteria freeze-dried powder, and 40-70% of solid fermentation culture of ganoderma lucidum bacteria, wherein the bacillus subtilis is CGMCC (China General Microbiological Culture Collection Center) 6012. The compound feed addictive disclosed by the invention can efficiently reduce the usage amount of antibiotic medicine in feeding animals, the safety of meat products of animals is improved, the utilization efficiency of the feed of the animals is improved, the immunity of the animals is improved, and the feeding efficiency is improved.
Description
Technical field:
The invention belongs to field of fodder, relate in particular to feed addictive.
Background technology:
In modern livestock and poultry cultivation process, in order to prevent and treat Animal diseases, often guarantee animal health with excessive antibiotic product, but excessive antibiotic use often causes the quality of animal meat product and product thereof to descend and the enhancing of antibiotic patience, and the transmission by food chain causes human health also to be subject to having a strong impact on of antibiotic problem.
Be derived from more natural natural materials and not only have good nutritive peculiarity, also have simultaneously antiviral, anti-inflammatory, anti-oxidant, regulate the effect such as body's immunity, have broad application prospects.
The main component of propolis is flavone compound, phenols, lactone, Coumarins, aldehyde ketone, the trace element such as a small amount of iron, calcium, silicon, manganese, lead, nickel, aluminium, copper, zinc and vitamin B complex, vitamin A, and several amino acids, enzyme, polysaccharide and abundant arsanilic acid etc.Propolis or domestic and international expert's " natural antibiotics " of generally acknowledging because it not only has the effect of the external invader of defence and infection pathogen diffusion, also have antiviral, anti-inflammatory, anti-oxidant, regulate the effect such as body's immunity.
The fruit of glossy privet claims again glossy privet reality, purpleflower holly fruit, Chinese wax tree, mouse Chinese catalpa.Exocarp is thin, and mesocarp is softer, easily peels off, and endocarp is wooden, yellowish-brown, and tool is indulged rib, breaks rear seed and is generally 1, kidney shape, atropurpureus, oiliness.Odorless, sweet, little bitterness of distinguishing the flavor of.Fruit of glossy privet fruit contains oleanolic acid (Oleanolic acid), sweet mellow wine, glucose, palmitic acid, stearic acid, oleic acid and leukotrienes.Pericarp contains ursolic acid (Ursolic acid), oleanolic acid, acetyloleanolic acid (Acetyloleanolic acid).Seed is fatty oily 14.9%, and in oil, palmitic acid and stearic acid are 19.5%, oleic acid and leukotrienes etc. be 80.5%.Still trace elements such as cupric, zinc, iron, manganese in the fruit of glossy privet.
Fruit of glossy privet tonifying liver kidney yin, crow must make eye bright, and modern medicine study thinks that the effect that the fruit of glossy privet can suppress helicobacter pylori can treat stomach trouble, also has and suppresses the treatment that the purine abnormal metabolism is used for gout and hyperuricemia.Be used for the deficiency of liver-yin and kidney-yin, soreness of waist tinnitus, poliosis; Unseeing, poor vision; Fever due to yin deficiency, stomach trouble and gout and and hyperuricemia.The fruit of glossy privet is clinical Chinese herbal medicine commonly used simply, and its effect in Production of Livestock and Poultry as feed addictive had in recent years begun one's study.
Antibacterial peptide is the biological small peptides that a class has antibacterial activity, is the important component part of natural animal immune defense system.The physicochemical property of antibacterial peptide is stable, and relative molecular mass is little, has a broad antifungal spectrum, and antibacterial mechanisms is unique.Compare with traditional antibiotic, have advantages of that antibacterial activity is strong, have no drug resistance, noresidue and toxicity little.In recent years, antibacterial peptide is the focus of research as antibiotic ideal substitute always.It is reported, antibacterial peptide has the generation that improves immune performance, reduces disease and the effect that improves survival rate as the additive feeding animals.Simultaneously, antibacterial peptide has good growth promoting function to animal, can improve the growth performance of animal, reduces feedstuff-meat ratio.Therefore, antibacterial peptide has the potentiality that become the green safety feed addictive.
Glossy ganoderma [Ganoderema lucidum (Leyss ex Fr.) karst] claims again polyporus lucidus, belongs to Basidiomycotina, Aphyllophorales, Ganodermataceae, Ganoderma.Doctor's allusion quotation in successive dynasties playbacks glossy ganoderma in " medicine-feeding ", and its ranking is before ginseng.The record of Shennong's Herbal, Compendium of Material Medica, glossy ganoderma materials for improving vision, tonifying liver gas, calm the nerves, beneficial essence, the beneficial motive, beneficial temper, beneficial lung qi, kidney-nourishing gas, logical nine orifices, ear is clever, order is bright, sharp joint, diuresis, skin maintenance, is first-class excellent tonic product.GL-B can promote the synthetic of protein, nucleic acid, and to the renewal of serum, liver and bone marrow cell protein or nucleic acid, synthetic facilitation arranged, it is one of glossy ganoderma main active ingredient of strengthening the body resistance to consolidate the constitution.GL-B mainly is present in lucid ganoderma fungus fructification, sclerotium, mycelium or zymotic fluid; Being present in ganoderic acid in glossy ganoderma pain relieving, calmness, inhibition histamine releasing are arranged, detoxify, protect the liver, kill the function of tumour cell, is one of main active ingredient of glossy ganoderma.
It is significant to the feed additive industry development how effective exploitation obtains the green feed additive product with natural plants or fungi.
Summary of the invention:
The invention provides the composite feed additive with antibiotic anti-oxidation function;
Composite feed additive is composed as follows:
Antibacterial peptide 10-15%, propolis 10-20%, fruit of glossy privet extract 10-25%, bacillus subtilis freeze-dried vaccine powder 15-30%, Ganoderma Lucidum solid fermentation culture 40-70%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on slant medium, cultivated about 96-120 hour, and covered with the inclined-plane to mycelia and get final product for 22-28 ℃; 25 ℃ of optimum culturing temperatures.
1. the liquid first order seed is cultivated: access of above-mentioned glossy ganoderma slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 22-28 ℃; 150 rev/mins of rotary shaking tables are advisable, and cultivation temperature is advisable with 25 ℃.
2. liquid two stage seed culture: the one-level shake-flask seed is equipped with the access of 5-20% inoculum concentration carries out secondary seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 22-28 ℃; Suitable inoculum concentration is 10%, and 150 rev/mins of rotary shaking tables are advisable, and cultivation temperature is advisable with 25 ℃.
3. solid fermentation is cultivated: the secondary shake-flask seed is equipped with in the 500L solid-state fermentation tank of the rear solid fermentation culture medium of sterilization with the access of 5-10% inoculum concentration, condition of culture: charge 50%, cultivation temperature 20-27 ℃, humidity 75-90%, ventilation 0.3-1.0 (V/V), incubation time 15-40 days.Opened every 3-10 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects the multiple drying means such as fluid bed, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The carbon source of solid fermentation culture medium of the present invention is mainly selected the raw materials commonly used such as wheat bran, glucose, sucrose, starch, corn flour, corncob (after pulverizing), and nitrogenous source can be selected groundnut meal, peptone, De-fatted soya protein Powder, beancake powder, dusty yeast etc.; The inorganic salts that need to add have potassium dihydrogen phosphate, magnesium sulfate.
Being added with mass ratio in above-mentioned solid fermentation culture medium is after 2-6% pulverizes, and after crossing the pulverizing of 40-50 purpose Radix Angelicae Sinensis, 3-8%, crosses 40-50 purpose Radix Isatidis, after 2-5% pulverizes, crosses 40-50 purpose thyme.
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and after crossing the 40-50 mesh sieve, adds 3-6 times of weight absolute ethyl alcohol dipping of the fruit of glossy privet and extracts, and controls 30-45 ℃ of temperature, adjust temperature after 2-4 hour and be 55-60 ℃ 1-2 hour, extract is concentrated, drying obtains ethanol extract; Add 75-85 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 2-4 times of fruit of glossy privet residue weight, processing time 30-50 minute, extract 2-3 time continuously, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
Described bacillus subtilis is bacillus subtilis CGMCC6012, and in the freeze-dried vaccine powder, number of viable is 4-8 * 108/g.Described composite feed additive is made by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract and bacillus subtilis freeze-dried vaccine powder proportionally are mixed to get composite feed additive.
The invention provides a kind of Feed Manufacturing bacillus subtilis (Bacillus subtilis) bacterial classification, this bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 16th, 2012, the preservation address is positioned at the Yard 1, BeiChen xi Road, Chaoyang District, Beijing City No. 3, (postcode 100101), deposit number are CGMCC No.6012.
Beneficial effect:
Bacillus subtilis: have following function: 1. oxygen consumption produces acid, and the regulating intestinal canal microecological balance reaches the diseases such as prevention diarrhoea, diarrhea; 2. secrete multiple enzyme, improve the feed digestive utilization ratio; 3. generation various nutrients participates in body metabolism; 4. raising animal immunizing power.
The fruit of glossy privet of the present invention carries out pulverize and break cellular wall to be processed, make in the fruit of glossy privet function factor realize full price stripping and high efficiency extraction utilization, adopt respectively organic solvent and hot water to realize wherein effective extraction of different efficacies composition, particularly control temperature section and extract extraction efficiency and the quality that has effectively improved especially glossy privet fruit extract.
Antibacterial peptide (antimicrobial peptides) is the general name with antibacterial activity small peptide.Antibacterial peptide has broad spectrum antibiotic activity, and bacterium is had very strong lethal effect, and especially it has more caused people's attention to the killing action of some drug resistance pathogen.In addition, people also find, some antibacterial peptide has killing action to part virus, fungi, protozoon and cancer cell etc., even can improve immunity, accelerating wound healing process.
The present invention is composite by science, bacillus subtilis, Ganoderma Lucidum have been realized organic assembling, particularly Ganoderma Lucidum has added traditional Chinese medicine ingredients in cultivating, product of the present invention can effectively reduce the use amount of antibiotics in letting animals feed, improve the security of animal meat product, improve the food utilization efficiency of animal, strengthen the immunity of animal, improve raise benefit.
Specific implementation method:
In the present invention, Ganoderma Lucidum culture production process is as follows:
(1) slant strains activation culture:
(2) the liquid first order seed is cultivated:
(3) liquid two stage seed culture:
(4) solid fermentation is cultivated:
(5) fermented and cultured finishes, and selects the multiple drying means such as fluid bed, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The present invention's lucidum strain used is purchased from Chinese industrial microorganism fungus kind preservation center, bacterium CICC14080.
In the present invention, the slant medium composition can be PDA slant medium or other suitable culture mediums.
The carbon source of solid fermentation culture medium of the present invention is mainly selected the raw materials commonly used such as wheat bran, glucose, sucrose, starch, corn flour, corncob (after pulverizing), and nitrogenous source can be selected groundnut meal, peptone, De-fatted soya protein Powder, beancake powder, dusty yeast etc.; The inorganic salts that need to add have potassium dihydrogen phosphate, magnesium sulfate.
Also being added with mass ratio in fermentation medium is the rear 40-50 of mistake of the pulverizing purpose Radix Isatidis of crossing 40-50 purpose Radix Angelicae Sinensis, 3-8% after 2-6% pulverizes, crosses 40-50 purpose thyme after 2-5% pulverizes;
In the present invention, the liquid fermentation seed culture medium forms: 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%, etiocobalamin Pm, PH6-7.
In the present invention, the glossy ganoderma cultural method is fit to belong to together other bacterial classification and carries out large scale fermentation by fermentation method and cultivate.
Induction mutation of bacterium
Take the ultraviolet mutagenesis method: take CICC10023 as starting strain, make bacteria suspension, coat on the culture medium plate, cultivate in 37 ℃ of incubators after ultraviolet irradiation 90s.
Bacterial screening
Choose 30 single bacterium colonies of looking strong from the 90s plate, be inoculated on the inclined-plane, in 37 ℃ of incubators, cultivated 24 hours, 0--4 ℃ stored refrigerated; With original strain in contrast, adopt the conventional herd breeding condition, sodium carboxymethylcellulose is as carbon source and proteolysis circle method screening cellulase-producing and the strong bacterial strain of protease, screening obtains cellulase-producing and strong bacterial strain bacillus subtilis (Bacillus subtilis) L1 of protease, this bacterial strain cellulase-producing can improve 25% by the force rate original strain, produces protease and can improve 52% by the force rate original strain.In the center preservation of Chinese common micro-organisms culture presevation administration committee's common micro-organisms, deposit number is CGMCC No 6012 with this bacterial classification.
This product bacillus subtilis used (Bacillus subtilis) bacterial classification, this bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on April 16th, 2012, the preservation address is positioned at the Yard 1, BeiChen xi Road, Chaoyang District, Beijing City No. 3, (postcode 100101), deposit number are CGMCC No6012.
Studies show that by experiment: product of the present invention have good inhibition Escherichia coli, staphylococcus aureus, etc. the characteristics of pathogenic bacteria.Adopt cylinder plate method that the fungistatic effect of ganoderma lucidum feedstuff additive product is studied, test adopts 10 times of water extracts of feed addictive to study.Experimental result sees Table 1, and result shows that product of the present invention has the superperformance that suppresses the pathogenic bacteria growth.
The fungistatic effect of table 1 feedstuff additive product
| Pathogenic bacteria | Staphylococcus aureus | Escherichia coli |
| Antibacterial circle diameter (mm) | 23 | 28 |
Safety is used in the stable performance of product of the present invention, with other feed addictive all without incompatibility.The addition of product of the present invention in feed is 1-2%, and it fully can the substitute antibiotics medicine, and noresidue, and is free from environmental pollution.After feeding, can significantly strengthen immunity and the premunition of animal body, promote growth, improve daily gain and feed conversion rate, and have anti-stress, antioxidant effect; Simultaneously intestinal bacteriosis there is stronger prevention effect.According to test, the aquatic products of feeding feed addictive of the present invention, quality improves, and meets the greenization production requirement of animal food, and Social benefit and economic benefit is very remarkable.
Product of the present invention is take Chinese traditional edible mushroom of biological fermentation process production as raw material, constant product quality is controlled, product noresidue in vivo, non-environmental-pollution, without the resistance to the action of a drug, and can improve immunity, have strong bacteriostasis, can improve culture benefit.Product meets the requirement of green feed additive fully.
Because glossy ganoderma has higher nutritive value and bacteria resistance function, can strengthen the immunity of animal, and also noresidue, environmentally safe.Producing glossy ganoderma and it is processed as feed addictive by biofermentation technique, meet the developing direction of current feed addictive fully, is a kind of green feed additive.
Embodiment 1(feedstuff additive product)
The invention provides the composite feed additive with antibiotic anti-oxidation function;
Composite feed additive is composed as follows:
Antibacterial peptide 10%, propolis 15%, fruit of glossy privet extract 15%, bacillus subtilis freeze-dried vaccine powder 20%, Ganoderma Lucidum solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
Antibacterial peptide is Shanghai Chu peptide bio tech ltd sell goods.
The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on slant medium, cultivated about 110 hours, and covered with the inclined-plane to mycelia and get final product for 25 ℃;
1. the liquid first order seed is cultivated: access of above-mentioned glossy ganoderma slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 100 rev/mins of rotary shaking tables, cultivated about 100 hours for 25 ℃; 25 ℃ of cultivation temperature.
2. liquid two stage seed culture: the one-level shake-flask seed is equipped with 10% inoculum concentration access carries out secondary seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 100 rev/mins of rotary shaking tables, cultivated about 130 hours for 25 ℃; 25 ℃ of cultivation temperature.
3. solid fermentation is cultivated: the secondary shake-flask seed is equipped with in the 500L solid-state fermentation tank of the rear solid fermentation culture medium of sterilization with 8% inoculum concentration access, condition of culture: charge 50%, 25 ℃ of cultivation temperature, humidity 85%, ventilation 0.5 (V/V), incubation time 25 days.Opened every 4 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects fluidized bed drying method, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
In the present invention, the solid fermentation culture medium is composed as follows: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 3% crosses 50 purpose Radix Angelicae Sinensis, 4% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 3% crosses 40 purpose thymes after pulverizing; The medium pH nature.Sterilising conditions: 121 ℃, 2 hours.
Described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized and is added 5 times of weight absolute ethyl alcohols dippings of the fruit of glossy privet after 45 mesh sieves and extract, and controls 35 ℃ of temperature, adjust temperature after 3 hours and be 57 ℃ 1.5 hours, extract is concentrated, drying obtains ethanol extract; Add 80 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 3 times of fruit of glossy privet residue weight, 40 minutes processing times, extract continuously 2 times, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
Described bacillus subtilis is bacillus subtilis CGMCC6012, and in the freeze-dried vaccine powder, number of viable is 6 * 10
8Individual/g.
Described composite feed additive is made by following methods:
After pulverizing, Ganoderma Lucidum culture, antibacterial peptide, propolis, fruit of glossy privet extract and bacillus subtilis freeze-dried vaccine powder proportionally are mixed to get composite feed additive.
Embodiment 2
The invention provides the composite feed additive with antibiotic anti-oxidation function;
Composed as follows:
Antibacterial peptide 15%, propolis 10%, fruit of glossy privet extract 25%, bacillus subtilis freeze-dried vaccine powder 10%, Ganoderma Lucidum solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on slant medium, cultivated about 110 hours, and covered with the inclined-plane to mycelia and get final product for 26 ℃;
1. the liquid first order seed is cultivated: access of above-mentioned glossy ganoderma slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 100 rev/mins of rotary shaking tables, cultivated about 100 hours for 26 ℃; 26 ℃ of cultivation temperature.
2. liquid two stage seed culture: the one-level shake-flask seed is equipped with 10% inoculum concentration access carries out secondary seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 100 rev/mins of rotary shaking tables, cultivated about 130 hours for 26 ℃; 26 ℃ of cultivation temperature.
3. solid fermentation is cultivated: the secondary shake-flask seed is equipped with in the 500L solid-state fermentation tank of the rear solid fermentation culture medium of sterilization with 8% inoculum concentration access, condition of culture: charge 50%, 26 ℃ of cultivation temperature, humidity 85%, ventilation 0.5 (V/V), incubation time 20 days.Opened every 3 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects fluidized bed drying method, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
In the present invention, the solid fermentation culture medium is composed as follows: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 3% crosses 50 purpose Radix Angelicae Sinensis, 5% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 2% crosses 40 purpose thymes after pulverizing;
Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.
Described fruit of glossy privet extract preparation method is as follows:
After the fruit of glossy privet was pulverized 50 mesh sieves, add 3 times of weight absolute ethyl alcohols dippings of the fruit of glossy privet and extract, control temperature 45 C, after 2 hours, adjust temperature and be 55 ℃ 1 hour, extract is concentrated, drying obtains ethanol extract; Add 85 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 4 times of fruit of glossy privet residue weight, 50 minutes processing times, extract continuously 3 times, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
Embodiment 3
Antibacterial peptide 10%, propolis 10%, fruit of glossy privet extract 10%, bacillus subtilis freeze-dried vaccine powder 10%, Ganoderma Lucidum solid fermentation culture 60%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on slant medium, cultivated about 120 hours, and covered with the inclined-plane to mycelia and get final product for 22 ℃; 25 ℃ of optimum culturing temperatures.
1. the liquid first order seed is cultivated: access of above-mentioned glossy ganoderma slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 80 rev/mins of rotary shaking tables, cultivated about 130 hours for 22 ℃;
2. liquid two stage seed culture: the one-level shake-flask seed is equipped with 5% inoculum concentration access carries out secondary seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 80 rev/mins of rotary shaking tables, cultivated about 96 hours for 22 ℃;
3. solid fermentation is cultivated: the secondary shake-flask seed is equipped with in the 500L solid-state fermentation tank of the rear solid fermentation culture medium of sterilization with 5% inoculum concentration access, condition of culture: charge 50%, 22 ℃ of cultivation temperature, humidity 75%, ventilation 0.3 (V/V), incubation time 15 days.Opened every 3 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects the multiple drying means such as fluid bed, and dry materials is arrived moisture 6%.The solid fermentation material is pulverized.
The solid fermentation culture medium is composed as follows: 70% wheat bran, 1.4% sucrose, 8% corn flour, 8% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; Culture medium PH nature.121 ℃ of sterilising conditions, 2 hours.4% crosses 50 purpose Radix Angelicae Sinensis, 3% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 3% crosses 40 purpose thymes after pulverizing;
Described fruit of glossy privet extract preparation method is as follows: the fruit of glossy privet is pulverized, and after crossing 50 mesh sieves, adds 6 times of weight absolute ethyl alcohols dippings of the fruit of glossy privet and extracts, and controls 30 ℃ of temperature, after 4 hours, adjust temperature and be 60 ℃ 2 hours, extract is concentrated, drying obtains ethanol extract; Add 85 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 2 times of fruit of glossy privet residue weight, 30 minutes processing times, extract continuously 2 times, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
Embodiment 4
Antibacterial peptide 12%, propolis 12%, fruit of glossy privet extract 14%, bacillus subtilis freeze-dried vaccine powder 13%, Ganoderma Lucidum solid fermentation culture 49%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on slant medium, cultivated about 96 hours, and covered with the inclined-plane to mycelia and get final product for 28 ℃;
1. the liquid first order seed is cultivated: access of above-mentioned glossy ganoderma slant strains picking is equipped with carries out first order seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 180 rev/mins of rotary shaking tables, cultivated about 96 hours for 28 ℃;
2. liquid two stage seed culture: the one-level shake-flask seed is equipped with 20% inoculum concentration access carries out secondary seed in 500 ml shake flasks of 100 milliliters of culture mediums and cultivate, condition of culture: 180 rev/mins of rotary shaking tables, cultivated about 96 hours for 28 ℃;
3. solid fermentation is cultivated: the secondary shake-flask seed is equipped with in the 500L solid-state fermentation tank of the rear solid fermentation culture medium of sterilization with 10% inoculum concentration access, condition of culture: charge 50%, 27 ℃ of cultivation temperature, humidity 90%, ventilation 1.0 (V/V), incubation time 40 days.Opened every 10 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects the multiple drying means such as fluid bed, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The solid fermentation culture medium is composed as follows: 66% wheat bran, 1.4% sucrose, 8% corn flour, 8% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.6% crosses 50 purpose Radix Angelicae Sinensis, 6% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 2% crosses 40 purpose thymes after pulverizing;
Described fruit of glossy privet extract preparation method is as follows:
After the fruit of glossy privet was pulverized the 40-50 mesh sieve, add 6 times of weight absolute ethyl alcohol dippings of the fruit of glossy privet and extract, control 35 ℃ of temperature, after 2 hours, adjusting temperature is 60 ℃, and 2 hours, extract was concentrated, drying obtains ethanol extract; Add 75 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 2 times of fruit of glossy privet residue weight, 50 minutes processing times, extract continuously 3 times, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
Embodiment 5 production methods are substantially with example 1
Antibacterial peptide 10%, propolis 12%, fruit of glossy privet extract 11%, bacillus subtilis freeze-dried vaccine powder 13%, Ganoderma Lucidum solid fermentation culture 54%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
The solid fermentation culture medium is composed as follows: 50% wheat bran, 1.4% sucrose, 10% corn flour, 21% corncob (after pulverizing), 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 2% crosses 50 purpose Radix Angelicae Sinensis, 8% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 5% crosses 40 purpose thymes after pulverizing; Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.
The product result of use
The result of use test of the inventive example 1 feed addictive in weanling pig
Test method:
It is 120 of the healthy weanling pigs of (7.16 ± 0.15) kg that experimental animal is selected Jing Zhou subordinate plant of twins feed corporation,Ltd average weight, and statistical analysis body weight difference is not remarkable.Adopt the single-factor Randomized Designs, with 120 healthy weanling pigs, by male and female half and half, be divided into 2 groups (control group and test group), every group of 6 repetitions, each repeats 10 pigs.Test group is added the product of the present invention that embodiment 1 makes, and control group does not add product of the present invention.Fed continuously 30 days, and compared with control group, use the invention feed addictive can obtain following effect:
| Project | Control group | Test group | Deviation |
| Starting weight (kg) | 7.09 | 7.23 | -- |
| End heavy (kg) | 18.34 | 19.95 | -- |
| Daily gain (g/d ﹒ head) | 375 | 424 | 8.7% |
| Daily ingestion amount (g/d ﹒ head) | 607.5 | 627.5 | 3.2% |
| Feedstuff-meat ratio | 1.62 | 1.48 | -8.6% |
| Diarrhea rate (%) | 14 | 2 | -85.7% |
Result of the test shows, adds this product in the weanling pig daily ration, and the daily gain of piglet improves a lot than control group; Feedstuff-meat ratio has obvious reduction; Diarrhea rate has reduced by 85.7%.
The result of use test of the inventive example 1 feed addictive in milking sow
Carried out the feeding experiment of 21 days by a definite date on the subordinate of twins feed corporation,Ltd pig farm, Jing Zhou.Single factor contrast design is adopted in test, chooses at random milking sow 30 health, that a farrowing number is close with the birth counterpoise, that parity is 2 or 3 tires, is divided at random 2 groups (being test group and control group), every group of 15 repetitions.Wherein: the test group daily ration adds this product that is prepared into by embodiment 2, and control group does not add product of the present invention.Record weaned piglet weight, diarrhea rate, the death rate.
| Project | Test group | Control group | Deviation |
| Produce the young number (number) of living | 10.9 | 11.0 | -- |
| Birth counterpoise (kg) | 1.46 | 1.48 | -- |
| 21 days small weaning pigs (head) | 10 | 8.30 | 20.4% |
| 21 days weight of weaning litters (kg) | 66.4 | 44.4 | 49.5% |
| 21 days wean counterpoises (kg) | 6.64 | 5.35 | 24.1% |
| Net gain (kg) | 5.18 | 3.87 | 33.8% |
| Daily gain (g) | 246 | 184 | 33.6% |
| Grice diarrhoea rate (%) | 6.1 | 14.3 | -57.3% |
| The death rate (%) | 1.6 | 5.4 | -70.3% |
Test shows: test group can improve 49.5% than control group weight of weaning litter, and a piglet daily gain improves 33.6%, and diarrhea rate reduces 57.3%, and the death rate reduces by 70.3%.
The above results shows that product of the present invention can improve sow and piglet body immunity, reduces antibiotic dosage, increases the cultivation quality and benefits.
Claims (6)
1. composite feed additive with antibiotic anti-oxidation function, described composite feed additive with antibiotic anti-oxidation function is composed as follows:
Antibacterial peptide 10-15%, propolis 10-20%, fruit of glossy privet extract 10-25%, bacillus subtilis freeze-dried vaccine powder 15-30%, Ganoderma Lucidum solid fermentation culture 40-70%, each constituent content sum is 100%; Bacillus subtilis (Bacillussubtilis) is CGMCC6012, and aforementioned proportion is mass percent; The preparation method is as follows for the Ganoderma Lucidum solid fermentation culture: the slant strains activation culture, the liquid first order seed is cultivated, the liquid two stage seed culture, solid fermentation is cultivated, fermented and cultured finishes, select fluidized bed drying method, dry materials is arrived moisture below 7%, the solid fermentation material is pulverized.
2. a kind of composite feed additive with antibiotic anti-oxidation function according to claim 1, it is characterized in that, being added with mass ratio in described Ganoderma Lucidum solid fermentation used medium is the rear 40-50 of mistake of the pulverizing purpose Radix Isatidis of crossing 40-50 purpose Radix Angelicae Sinensis, 3-8% after 2-6% pulverizes, crosses 40-50 purpose thyme after 2-5% pulverizes.
3. a kind of composite feed additive with antibiotic anti-oxidation function according to claim 1, is characterized in that, described fruit of glossy privet extract preparation method is as follows:
The fruit of glossy privet is pulverized, and after crossing the 40-50 mesh sieve, adds 3-6 times of weight absolute ethyl alcohol dipping of the fruit of glossy privet and extracts, and controls 30-45 ℃ of temperature, after 2-4 hour, adjust temperature and be 55-60 ℃ 1-2 hour, extract is concentrated, drying obtains ethanol extract; Add 75--85 ℃ of hot water in fruit of glossy privet residue after alcohol extract, the hot water addition is 2-4 times of fruit of glossy privet residue weight, processing time 30-50 minute, extract 2-3 time continuously, and with spray-drying after the extract Vacuum Concentration, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, namely get glossy privet fruit extract.
4. a kind of composite feed additive with antibiotic anti-oxidation function according to claim 1, is characterized in that, in described bacillus subtilis freeze-dried vaccine powder, number of viable is 4-8 * 10
8Individual/g.
5. a kind of composite feed additive with antibiotic anti-oxidation function according to claim 1, is characterized in that, described composite feed additive with antibiotic anti-oxidation function is composed as follows:
Antibacterial peptide 10%, propolis 15%, fruit of glossy privet extract 15%, bacillus subtilis freeze-dried vaccine powder 20%, Ganoderma Lucidum solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is mass percent.
6. a kind of composite feed additive with antibiotic anti-oxidation function according to claim 1, it is characterized in that, described Ganoderma Lucidum solid fermentation culture medium is composed as follows: the corncob after 70% wheat bran, 1.4% sucrose, 8% corn flour, 8% are pulverized, 0.1% peptone, 2% De-fatted soya protein Powder, 0.5% dusty yeast; 4% crosses 50 purpose Radix Angelicae Sinensis, 3% rear mistake 40 purpose Radix Isatidis of pulverizing after pulverizing, and 3% crosses 40 purpose thymes after pulverizing.
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| CN104366020A (en) * | 2014-09-28 | 2015-02-25 | 天津天绿健科技有限公司 | Feed additive product |
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| CN104718990B (en) * | 2015-03-27 | 2015-11-11 | 宾石其 | The production method of a kind of composite fungi feed production and Edible Fungi unification |
| CN105028954A (en) * | 2015-08-26 | 2015-11-11 | 天津天绿健科技有限公司 | Feed additive product |
| CN105146103A (en) * | 2015-09-17 | 2015-12-16 | 天津中天精科科技有限公司 | Feed additive |
| CN105166410A (en) * | 2015-09-17 | 2015-12-23 | 邵素英 | Feed additive with good thermal stability |
| CN105231023A (en) * | 2015-09-17 | 2016-01-13 | 邵素英 | Compound feed additive with good thermal stability |
| CN105192355A (en) * | 2015-09-17 | 2015-12-30 | 邵素英 | Feed additive rich in carotene |
| CN105146104A (en) * | 2015-09-17 | 2015-12-16 | 天津天绿健科技有限公司 | Feed additive containing rhodotorula |
| CN105146093A (en) * | 2015-09-17 | 2015-12-16 | 天津中天精科科技有限公司 | Compound feed additive rich in carotenes |
| CN105192311A (en) * | 2015-09-17 | 2015-12-30 | 邵素英 | Livestock and poultry compound feed additive |
| CN105192310A (en) * | 2015-09-17 | 2015-12-30 | 天津天绿健科技有限公司 | Compound feed additive containing rhodotorula |
| CN105104758A (en) * | 2015-09-17 | 2015-12-02 | 北京科慧通智慧科技有限公司 | Livestock and poultry feed additive |
| CN105192309A (en) * | 2015-09-17 | 2015-12-30 | 天津中天精科科技有限公司 | Composite feed additive |
| CN105166323B (en) * | 2015-09-25 | 2019-01-22 | 广东海洋大学 | A kind of composite feed additive |
| CN106879823A (en) * | 2016-12-31 | 2017-06-23 | 新昌县田野泉养殖技术开发有限公司 | The composite feed additive of antibacterial anti-oxidation function |
| CN109874909A (en) * | 2019-03-28 | 2019-06-14 | 陈欣 | A kind of feed addictive and preparation process based on antibacterial peptide |
| CN111607627A (en) * | 2020-05-12 | 2020-09-01 | 南京大方生物工程有限公司 | Fermentation process for producing antibacterial peptide bacillus subtilis and application of fermentation process in disease-resistant and yield-increasing of livestock and poultry |
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