CN102657287A - 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 the modern livestock and poultry cultivation process; In order to prevent and treat Animal diseases; The excessive antibiotic product of normal use is guaranteed animal health; But excessive antibiotic use often causes the quality of animal flesh goods and product thereof to descend and the enhancing of antibiotic patience, and the transmission through food chain causes human health also to receive 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 effect such as body's immunity, have broad application prospects.
The main component of propolis is flavone compound, phenols, lactone, Coumarins, aldehyde ketone; Trace element such as a spot 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 still is " the natural antibiotics " that domestic and international expert generally acknowledges because it not only has the effect of external invador of defence and infection pathogen diffusion, also have antiviral, anti-inflammatory, anti-oxidant, regulate effect such as body's immunity.
The fruit of glossy privet is claimed glossy privet reality, purpleflower holly fruit, Chinese wax tree, mouse Chinese catalpa again.Exocarp is thin, and mesocarp is softer, is prone to peel off, and endocarp is wooden, yellowish-brown, and tool is indulged rib, breaks the back 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 palmitic acid and stearic acid are 19.5% in the oil, 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 simply Chinese herbal medicine commonly used, has begun one's study it in recent years as the effect of feed addictive in Production of Livestock and Poultry.
Antibacterial peptide is one type of biological small peptides with 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 antibacterial activity strong, have no drug resistance, noresidue and the little advantage of toxicity.In recent years, antibacterial peptide is the focus of research as antibiotic ideal substitute always.It is reported that antibacterial peptide has 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] is claimed polyporus lucidus again, belongs to Basidiomycotina, Aphyllophorales, Ganodermataceae, Ganoderma.Doctor's allusion quotation in successive dynasties playbacks glossy ganoderma in " medicine-feeding ", and its ranking is before genseng.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 acute hearing, bright, the sharp joint of order, diuresis, skin maintenance, be 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 the zymotic fluid; The ganoderic acid that is present in the glossy ganoderma has pain relieving, calmness, inhibition histamine to discharge, detoxify, protect the liver, kill the function of tumour cell, is one of main active ingredient of glossy ganoderma.
How effectively exploitation is significant to the feed additive industry development with natural plants or fungi acquisition green feed additive product.
Summary of the invention:
The present invention provides the composite feed additive with antibiotic anti-oxidation function;
Composite feed additive is formed as follows:
Antibacterial peptide 10-15%, propolis 10-20%, fruit of glossy privet extract 10-25%, bacillus subtilis freeze-dried vaccine powder 15-30%, glossy ganoderma bacteria solid fermentation culture 40-70%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
The preparation method is following for glossy ganoderma bacteria solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on the 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 ℃; Rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 25 ℃.
2. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with the 5-20% inoculum concentration carry out the secondary seed cultivation in 500 ml shake flasks that 100 milliliters of culture mediums are housed, condition of culture: rotary shaking table 80-180 rev/min, cultivated about 96-130 hour for 22-28 ℃; Suitable inoculum concentration is 10%, and rotary shaking table is advisable for 150 rev/mins, and cultivation temperature is advisable with 25 ℃.
3. solid fermentation is cultivated: insert the secondary shake-flask seed in the 500L solid-state fermentation tank that sterilization back solid fermentation culture medium is housed with the 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.The every unlatching at a distance from 3-10 days stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects 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 wheat bran, glucose, sucrose, starch, corn flour, corncob raw materials commonly used such as (pulverizing the back), and nitrogenous source can be selected groundnut meal, peptone, skimmed soybean protein powder, soya-bean cake powder, dusty yeast etc.; The inorganic salts that need to add have potassium dihydrogen phosphate, magnesium sulfate.
In above-mentioned solid fermentation culture medium, be added with mass ratio and be after 2-6% pulverizes, cross the pulverizing of 40-50 purpose Radix Angelicae Sinensis, 3-8% after, cross 40-50 purpose Radix Isatidis, after 2-5% pulverizes, cross 40-50 purpose thyme.
Said fruit of glossy privet extract preparation method is following:
The fruit of glossy privet is pulverized, cross the 40-50 mesh sieve after, add 3-6 times of weight absolute ethyl alcohol dipping of the fruit of glossy privet and extract, 30-45 ℃ of control temperature, after 2-4 hour the adjustment temperature be 55-60 ℃ 1-2 hour, extract concentrates, drying obtains ethanol extract; Add 75-85 ℃ of hot water in the fruit of glossy privet residue behind the alcohol extract, the hot water addition is 2-4 a times of fruit of glossy privet residue weight, processing time 30-50 minute, extract 2-3 time continuously, and the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly get glossy privet fruit extract.
Said bacillus subtilis is bacillus subtilis CGMCC6012, and number of viable is 4-8 * 108/g in the freeze-dried vaccine powder.Said composite feed additive is made by following method:
Pulverizing back glossy ganoderma bacterium culture, antibacterial peptide, propolis, fruit of glossy privet extract and bacillus subtilis freeze-dried vaccine powder proportionally mixes and obtains composite feed additive.
The present invention provides a kind of Feed Manufacturing with bacillus subtilis (Bacillus subtilis) bacterial classification; This bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms 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, regulates the intestinal microecology balance, reaches diseases such as prevention diarrhoea, diarrhea; 2. secrete multiple enzyme, improve the feed digestive utilization ratio; 3. generation various nutrients is participated 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 handled; Make function factor realization full price stripping and high efficiency extraction utilization in the fruit of glossy privet; Adopt organic solvent and hot water to realize wherein effective extraction of different efficacies composition respectively, particularly control temperature section and extract extraction efficiency and the quality that has effectively improved glossy privet fruit extract especially.
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 some drug resistance kill pathogenic bacteria effect.In addition, people find that also 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 through science; Bacillus subtilis, glossy ganoderma bacterium have been realized organic assembling, and particularly the glossy ganoderma bacterium has been added traditional Chinese medicine ingredients in cultivating, and product of the present invention can effectively reduce the use amount of antibiotics in letting animals feed; Improve the security of animal flesh goods; Improve the food utilization efficiency of animal, strengthen the immunity of animal, improve raise benefit.
The practical implementation method:
Glossy ganoderma bacterium culture production process is following among the present invention:
(1) slant strains activation culture:
(2) the liquid first order seed is cultivated:
(3) the liquid secondary seed is cultivated:
(4) solid fermentation is cultivated:
(5) fermented and cultured finishes, and selects multiple drying means such as fluid bed, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The used lucidum strain of the present invention is purchased in Chinese industrial microorganism fungus kind preservation center, bacterium CICC14080.
The slant medium composition can be PDA slant medium or other suitable culture mediums among the present invention.
The carbon source of solid fermentation culture medium of the present invention is mainly selected wheat bran, glucose, sucrose, starch, corn flour, corncob raw materials commonly used such as (pulverizing the back), and nitrogenous source can be selected groundnut meal, peptone, skimmed soybean protein powder, soya-bean cake powder, dusty yeast etc.; The inorganic salts that need to add have potassium dihydrogen phosphate, magnesium sulfate.
In fermentation medium, also be added with mass ratio and be 2-6% and pulverize and cross 40-50 purpose Radix Isatidis after the pulverizing of 40-50 purpose Radix Angelicae Sinensis, 3-8% is crossed in the back, 2-5% pulverizes the back and crosses 40-50 purpose thyme;
The liquid fermentation seed culture medium is formed among the present invention: starch 2-3%, sucrose 3-5%, glucose 1-3%, peptone 0.5%, skimmed soybean protein powder 2-3%, potassium dihydrogen phosphate 0.13-0.2%, magnesium sulfate 0.1-0.15%, etiocobalamin Pm, PH6-7.
The glossy ganoderma cultural method is fit to belong to together other bacterial classification and carries out large scale fermentation through fermentation method and cultivate among the present invention.
Induction mutation of bacterium
Take the ultraviolet mutagenesis method: with CICC10023 is starting strain, processes bacteria suspension, coats on the culture medium plate, cultivates in 37 ℃ of incubators behind the 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 0--4 ℃ of stored refrigerated 24 hours; With original strain as contrast; Adopt conventional seed selection 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, and 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 common micro-organisms center preservation of Chinese common micro-organisms culture presevation administration committee, deposit number is CGMCC No 6012 with this bacterial classification.
The used bacillus subtilis of this product (Bacillus subtilis) bacterial classification; This bacterial strain has been preserved in China Committee for Culture Collection of Microorganisms 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.
Show through experimental study: 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 is seen table 1, and the 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 |
Performance of products of the present invention is stable, and is safe in utilization, all do not have incompatibility with other feed addictive.The addition of product of the present invention in feed is 1-2%, and it can substitute antibiotic medicine fully, and noresidue, and is free from environmental pollution.After feeding, can significantly strengthen the 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 are very significantly.
Product of the present invention is a raw material with Chinese traditional edible mushroom of biological fermentation process production; Constant product quality is controlled; Product noresidue in vivo, non-environmental-pollution, the no resistance to the action of a drug, and can improve immunity, have strong bacteriostasis, culture benefit can be improved.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 noresidue, environmentally safe.Producing glossy ganoderma and it is processed as feed addictive through biofermentation technique, meet the developing direction of current feed addictive fully, is a kind of green feed additive.
Embodiment 1 (feedstuff additive product)
The present invention provides the composite feed additive with antibiotic anti-oxidation function;
Composite feed additive is formed as follows:
Antibacterial peptide 10%, propolis 15%, fruit of glossy privet extract 15%, bacillus subtilis freeze-dried vaccine powder 20%, glossy ganoderma bacteria solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
Antibacterial peptide is Shanghai Chu peptide bio tech ltd sell goods.
The preparation method is following for glossy ganoderma bacteria solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on the 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. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 10% inoculum concentration carry out the secondary seed cultivation in 500 ml shake flasks that 100 milliliters of culture mediums are housed, 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 inserted in the 500L solid-state fermentation tank that sterilization back solid fermentation culture medium is housed condition of culture: charge 50%, 25 ℃ of cultivation temperature, humidity 85%, ventilation 0.5 (V/V), incubation time 25 days with 8% inoculum concentration.Whenever opened at a distance from 4 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects the fluidized bed drying method, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The solid fermentation culture medium is formed as follows among the present invention: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; 3% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 4% pulverizing are crossed in the back, and 3% pulverizes the back crosses 40 purpose thymes; The medium pH nature.Sterilising conditions: 121 ℃, 2 hours.
Said fruit of glossy privet extract preparation method is following:
The fruit of glossy privet is pulverized and is added 5 times of weight absolute ethyl alcohols dippings of the fruit of glossy privet behind 45 mesh sieves and extract, 35 ℃ of control temperature, after 3 hours the adjustment temperature be 57 ℃ 1.5 hours, extract concentrates, drying obtains ethanol extract; Add 80 ℃ of hot water in the fruit of glossy privet residue behind the 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 the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly get glossy privet fruit extract.
Said bacillus subtilis is bacillus subtilis CGMCC6012, and number of viable is 6 * 10 in the freeze-dried vaccine powder
8Individual/g.
Said composite feed additive is made by following method:
Pulverizing back glossy ganoderma bacterium culture, antibacterial peptide, propolis, fruit of glossy privet extract and bacillus subtilis freeze-dried vaccine powder proportionally mixes and obtains composite feed additive.
Embodiment 2
The present invention provides the composite feed additive with antibiotic anti-oxidation function;
Form as follows:
Antibacterial peptide 15%, propolis 10%, fruit of glossy privet extract 25%, bacillus subtilis freeze-dried vaccine powder 10%, glossy ganoderma bacteria solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
The preparation method is following for glossy ganoderma bacteria solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on the 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. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 10% inoculum concentration carry out the secondary seed cultivation in 500 ml shake flasks that 100 milliliters of culture mediums are housed, 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 inserted in the 500L solid-state fermentation tank that sterilization back solid fermentation culture medium is housed condition of culture: charge 50%, 26 ℃ of cultivation temperature, humidity 85%, ventilation 0.5 (V/V), incubation time 20 days with 8% inoculum concentration.Whenever opened at a distance from 3 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects the fluidized bed drying method, and dry materials is arrived moisture below 7%.The solid fermentation material is pulverized.
The solid fermentation culture medium is formed as follows among the present invention: 60% wheat bran, 1.4% sucrose, 10% corn flour, 16% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; 3% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 5% pulverizing are crossed in the back, and 2% pulverizes the back crosses 40 purpose thymes;
Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.
Said fruit of glossy privet extract preparation method is following:
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, 45 ℃ of control temperature, after 2 hours, the adjustment temperature be 55 ℃ 1 hour, extract concentrates, drying obtains ethanol extract; Add 85 ℃ of hot water in the fruit of glossy privet residue behind the 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 the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly 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%, glossy ganoderma bacteria solid fermentation culture 60%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
The preparation method is following for glossy ganoderma bacteria solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on the 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. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 5% inoculum concentration carry out the secondary seed cultivation in 500 ml shake flasks that 100 milliliters of culture mediums are housed, 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 inserted in the 500L solid-state fermentation tank that sterilization back solid fermentation culture medium is housed condition of culture: charge 50%, 22 ℃ of cultivation temperature, humidity 75%, ventilation 0.3 (V/V), incubation time 15 days with 5% inoculum concentration.Whenever opened at a distance from 3 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects 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 formed as follows: 70% wheat bran, 1.4% sucrose, 8% corn flour, 8% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; Culture medium PH nature.121 ℃ of sterilising conditions, 2 hours.4% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 3% pulverizing are crossed in the back, and 3% pulverizes the back crosses 40 purpose thymes;
Said fruit of glossy privet extract preparation method is following: the fruit of glossy privet is pulverized, cross 50 mesh sieves after, add 6 times of weight absolute ethyl alcohols dippings of the fruit of glossy privet and extract, 30 ℃ of control temperature, after 4 hours, the adjustment temperature be 60 ℃ 2 hours, extract concentrates, drying obtains ethanol extract; Add 85 ℃ of hot water in the fruit of glossy privet residue behind the 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 the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly 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%, glossy ganoderma bacteria solid fermentation culture 49%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
The preparation method is following for glossy ganoderma bacteria solid fermentation culture:
Slant strains activation culture: the glossy ganoderma slant strains of preservation is transferred on the 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. the liquid secondary seed is cultivated: the one-level shake-flask seed is inserted with 20% inoculum concentration carry out the secondary seed cultivation in 500 ml shake flasks that 100 milliliters of culture mediums are housed, 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 inserted in the 500L solid-state fermentation tank that sterilization back solid fermentation culture medium is housed condition of culture: charge 50%, 27 ℃ of cultivation temperature, humidity 90%, ventilation 1.0 (V/V), incubation time 40 days with 10% inoculum concentration.Whenever opened at a distance from 10 days and stirred 10 minutes.Adopt the SGF solid-state fermentation tank.
4. fermented and cultured finishes, and selects 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 formed as follows: 66% wheat bran, 1.4% sucrose, 8% corn flour, 8% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.6% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 6% pulverizing are crossed in the back, and 2% pulverizes the back crosses 40 purpose thymes;
Said fruit of glossy privet extract preparation method is following:
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, 35 ℃ of control temperature, after 2 hours, the adjustment temperature is 60 ℃, 2 hours, extract concentrated, drying obtains ethanol extract; Add 75 ℃ of hot water in the fruit of glossy privet residue behind the 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 the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly get glossy privet fruit extract.
Embodiment 5 production methods are basic with example 1
Antibacterial peptide 10%, propolis 12%, fruit of glossy privet extract 11%, bacillus subtilis freeze-dried vaccine powder 13%, glossy ganoderma bacteria solid fermentation culture 54%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
The solid fermentation culture medium is formed as follows: 50% wheat bran, 1.4% sucrose, 10% corn flour, 21% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; 2% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 8% pulverizing are crossed in the back, and 5% pulverizes the back crosses 40 purpose thymes; Culture medium PH nature.Sterilising conditions, 121 ℃, 2 hours.
The product result of use
The result of use test of the present invention's example 1 feed addictive in weanling pig
Test method:
Experimental animal selects Jing Zhou subordinate plant of twins feed corporation,Ltd average weight to be 120 of the healthy weanling pigs of (7.16 ± 0.15) kg, and is not remarkable through statistical analysis body weight difference.Adopt the single-factor Randomized Designs,,, be divided into 2 groups (control group and test group) by public mother half and half with 120 healthy weanling pigs, 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, use the invention feed addictive can obtain following effect with control group:
| 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, in the weanling pig daily ration, adds this product, 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 present invention's example 1 feed addictive in milking sow
Twins feed corporation,Ltd has carried out on the subordinate pig farm 21 days feeding experiment by a definite date in Jing Zhou.Single factor contrast design is adopted in test, and the milking sow 30 health of picked at random, that a farrowing number is close with the birth counterpoise, that parity is 2 or 3 tires is divided into 2 groups (being test group and control group), every group of 15 repetitions at random.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 to culture quality and benefits.
Claims (9)
1. composite feed additive with antibiotic anti-oxidation function, said composite feed additive with antibiotic anti-oxidation function is formed as follows:
Antibacterial peptide 10-15%, propolis 10-20%, fruit of glossy privet extract 10-25%, bacillus subtilis freeze-dried vaccine powder 15-30%, glossy ganoderma bacteria solid fermentation culture 40-70%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent; The preparation method is following for glossy ganoderma bacteria solid fermentation culture: the slant strains activation culture; The liquid first order seed is cultivated, and the liquid secondary seed is cultivated, and solid fermentation is cultivated; Fermented and cultured finishes; Select the fluidized bed drying method, dry materials is arrived moisture below 7%, the solid fermentation material is pulverized.
2. according to the said a kind of composite feed additive of claim 1 with antibiotic anti-oxidation function; It is characterized in that; Be added with mass ratio in the used culture medium of said glossy ganoderma bacteria solid fermentation and be 2-6% and pulverize and cross 40-50 purpose Radix Isatidis after the pulverizing of 40-50 purpose Radix Angelicae Sinensis, 3-8% is crossed in the back, 2-5% pulverizes the back and crosses 40-50 purpose thyme.
3. according to the said a kind of composite feed additive of claim 1, it is characterized in that said fruit of glossy privet extract preparation method is following with antibiotic anti-oxidation function:
The fruit of glossy privet is pulverized, cross the 40-50 mesh sieve after, add 3-6 times of weight absolute ethyl alcohol dipping of the fruit of glossy privet and extract, 30-45 ℃ of control temperature, after 2-4 hour, the adjustment temperature be 55-60 ℃ 1-2 hour, extract concentrates, drying obtains ethanol extract; Add 75--85 ℃ of hot water in the fruit of glossy privet residue behind the alcohol extract, the hot water addition is 2-4 a times of fruit of glossy privet residue weight, processing time 30-50 minute, extract 2-3 time continuously, and the extract vacuum is concentrated the back spray-drying, obtain the hot water extract; Above-mentioned ethanol extract and hot water extract are merged pulverizing, cross 60 mesh sieves, promptly get glossy privet fruit extract.
4. according to the said a kind of composite feed additive of claim 1, it is characterized in that number of viable is 4-8 * 10 in the said bacillus subtilis freeze-dried vaccine powder with antibiotic anti-oxidation function
8Individual/g.
5. according to the said a kind of composite feed additive of claim 1, it is characterized in that said composite feed additive with antibiotic anti-oxidation function is formed as follows with antibiotic anti-oxidation function:
Antibacterial peptide 10%, propolis 15%, fruit of glossy privet extract 15%, bacillus subtilis freeze-dried vaccine powder 20%, glossy ganoderma bacteria solid fermentation culture 40%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
6. according to the said a kind of composite feed additive of claim 1, it is characterized in that said composite feed additive with antibiotic anti-oxidation function is formed as follows with antibiotic anti-oxidation function:
Antibacterial peptide 10%, propolis 10%, fruit of glossy privet extract 10%, bacillus subtilis freeze-dried vaccine powder 10%, glossy ganoderma bacteria solid fermentation culture 60%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
7. according to the said a kind of composite feed additive of claim 1, it is characterized in that said composite feed additive with antibiotic anti-oxidation function is formed as follows with antibiotic anti-oxidation function:
Antibacterial peptide 12%, propolis 12%, fruit of glossy privet extract 14%, bacillus subtilis freeze-dried vaccine powder 13%, glossy ganoderma bacteria solid fermentation culture 49%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
8. according to the said a kind of composite feed additive of claim 1 with antibiotic anti-oxidation function; It is characterized in that; Said glossy ganoderma bacteria solid fermentation culture medium is formed as follows: 70% wheat bran, 1.4% sucrose, 8% corn flour, 8% corncob (pulverizing the back), 0.1% peptone, 2% skimmed soybean protein powder, 0.5% dusty yeast; 4% pulverizes and to cross 40 purpose Radix Isatidis after 50 purpose Radix Angelicae Sinensis, 3% pulverizing are crossed in the back, and 3% pulverizes the back crosses 40 purpose thymes.
9. according to the said a kind of composite feed additive of claim 1, it is characterized in that said composite feed additive with antibiotic anti-oxidation function is formed as follows with antibiotic anti-oxidation function:
Antibacterial peptide 10%, propolis 12%, fruit of glossy privet extract 11%, bacillus subtilis freeze-dried vaccine powder 13%, glossy ganoderma bacteria solid fermentation culture 54%; Bacillus subtilis (Bacillus subtilis) is CGMCC6012, and aforementioned proportion is a mass percent.
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Application publication date: 20120912 Assignee: QINGYUAN JIANGFENG FERTILIZER CO., LTD. Assignor: Jingzhou Twins Feed Co., Ltd. Contract record no.: 2015120000024 Denomination of invention: Compound feed addictive with bacteria and oxidation resisting functions Granted publication date: 20130508 License type: Exclusive License Record date: 20150513 |
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