CN1965673A - Micro-ecological preparation - Google Patents
Micro-ecological preparation Download PDFInfo
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- CN1965673A CN1965673A CNA2005101104865A CN200510110486A CN1965673A CN 1965673 A CN1965673 A CN 1965673A CN A2005101104865 A CNA2005101104865 A CN A2005101104865A CN 200510110486 A CN200510110486 A CN 200510110486A CN 1965673 A CN1965673 A CN 1965673A
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Abstract
The invention relates to a method for using mixture mushroom to cultivate the microecosystem agent with microbe viable bacteria agent and enzyme agent, wherein said agent is formed by one or more of lactic acid bacillus, bacillus, saccharomycete, filamentous fungi and photosynthetic bacteria, to be cultivated in cultivate substrate, while the ferment temperature is 28-32Deg. C and the ferment time is 40-50h, to obtain the microecosystem agent. The invention can adjust the microecosystem balance; accelerate the adsorption mechanism of intestinal canal, etc.
Description
Technical field
The present invention relates to microorganism field, more specifically reference and a kind of prebiotics feed additive.It can replenish the beneficial microbe that reduces or lack in the animal intestinal on quantity or kind, adjust or keep the microecological balance in the enteron aisle, strengthens human body immune function and anti-stress ability, improves culture benefit.
Background technology
Usually single microorganism produces microbe metabolite through fermentation, utilize this metabolin product to cure the disease as antibiotic, the utility of feed interpolation or the like, for example: Chinese patent CN 200410077706.4, microbial fermentation is passed through in this invention, additive adds in the feed or preparation non-antibiotic class feed, can significantly improve the animal immune ability, promote the metabolism of animal, improve the resistance against diseases of animal body, improve the digestive function of animal, promote growth of animal, and obviously improve the meat of animal.Application and development trend at microorganism formulation, it is kind of a natural biologically active agents that this article has been summarized probiotics-probiotic, have no side effect, have no drug resistance, the drug residue free thing, have effect (sources: Shandong poultry 2001, (5): the 31-32 page or leaf) such as health care, promotion growth, raising efficiency of feed utilization.United States Patent (USP) relates to a kind of method that detects probiotics quality standard in the animal feed.Contain micro-ecological bacterial and oligosaccharides in this feed, detection method is mainly hybridization and polymerase chain reaction technology (TI:Methods for detecting and quantifying specificprobiotic microorganisms in animal feed.Inventor:amer BE.Application number:S2004000711155 Patent number:S20050048515).
Summary of the invention
The purpose of this invention is to provide a kind of probiotics that adopts Mixed Microbes to cultivate to have microorganism live bacteria preparation and enzyme preparation double effects, produce in order to overcome probiotic on the present domestic market that ubiquitous kind, effect are single, the problem of poor stability.
For probiotics, guarantee the normal performance of rendeing a service of probio, just should guarantee content and the stability of viable bacteria in the active bacteria formulation, make it can stand the influence of extraneous factor.This novel complex micro organism fungicide is with reference to the international research main flow direction, the probio of single bacterial strain is converted into composite bacteria agent capable, therefore has multiple efficacies, it not only can regulate the microecological balance of body, promote the absorption of enteron aisle, and the microorganism in the said preparation can also secrete useful nutritional labelings such as multivitamin and amino acid, improve the body function, and produce the organic acid antibiotic, kill pathogenic bacteria is prevented and cured diseases.The enzyme of the amino acid oxidase of its generation and branch system sulfide, but harmful substances such as cracked ammonium, hydrogen sulfide, indoles reduce content of harmful gas in the hen house, reduce stink, improve environment, minimizing chicken group's the incidence of disease.
In addition, advanced microcapsule embedded technology has been adopted in the production of probiotics of the present invention, makes wherein microorganism can stand the influence of external conditions such as temperature, humidity, acidity, has guaranteed that most of probios produce more scabrous stability problem.
Finished product of the present invention is the beneficial microbe that separates in animal body, grow through special process fermentation training and to make, only contain viable bacteria or comprise thalline and the active bacteria formulation of metabolite, one or more bacterium of Bacillus acidi lactici, bacillus, saccharomycete, filamentous fungi and photosynthetic bacteria are formed.Technological process is: bacterial screening → bacterial classification cultivation → raw material → liquid fermentation → post processing → finished product
1. bacterial screening: when selecting to produce bacterial strain,, except using traditional fermentation strain, screening other bacterial classifications of using must meet following condition: 1. can assimilate matrix carbon source and inorganic nitrogen-sourced well; 2. reproduction speed is fast, mycoprotein content height; 3. avirulence and pathogenic; 4. bacterial classification stable performance; 5. preferably can mix bacterium cultivates.
2. bacterial classification training is grown: adopt many bacterial classifications to mix bacterium and cultivate to require to exist between bacterial strain and live together or laterality is given birth to or symbiosis, we adopt good bacterium by solid fermentation crude starch material and slag cypress class raw material.
3. raw material: according to the growth requirement of bacterial classification, from domestic tens kinds of main agricultural byproducts through fermentation relatively culture medium be that main material adds that a small amount of assistant agent and trace element form by analysis for soybean powder, corn, rice bran, dregs of beans.
4. fermentation and processing: aspect production technology, be to guarantee product quality, adopt liquid fermentation process, thereby improve the controllability and the production scale of producing, reduce production costs.The technology of preparing of this probiotics is by filamentous fungi, saccharomycete, bacillus, lactic acid bacteria, compositions such as photosynthetic bacteria.Preceding four kinds of bacterium respectively under 28-32 ℃ of temperature environment, solid or liquid fermentation 40-50 hour; Wherein lactic acid bacteria was by 1: 1 adding inserts; In 35-50 ℃ of environment, dry, pulverize, sieve through the 40-60 order; Photosynthetic bacteria carries out seeing under 20 ℃ of environment of light; Add minerals and vitamins, mix and make.
5. finished product
Classificating requirement
Table 1 technical target of the product
The innoxious index of table 2 finished product
| Project | Index |
| Salmonella, individual/ml≤ | Must not detect |
| Mould, individual/ml≤ | Must not detect |
| Coliform, individual/ml≤ | 0.3 |
| Mercury and compound (in Hg), mg/kg≤ | 0.05 |
| Cadmium and compound (in Cd), mg/kg≤ | 0.2 |
| Arsenic and compound (in As), mg/kg≤ | 0.5 |
| Lead and compound (in Pb), mg/kg≤ | 1 |
Beneficial effect
Standing grain dimension probiotic is a kind of powerful microorganism species, after plant uses it, plant's stink is significantly reduced or disappear, efficiency of feed utilization increases substantially, the poultry egg production increases, the premunition of animal strengthens, breeding potential improves, and can replace with growth promotion, pre-disease is purpose medicated premix, enzyme preparation etc., thereby produces the green feed and the cost of manufacture that reduces feed of drug residue free.And the animal product of producing is a pollution-free food.
It has the double effects of microorganism live bacteria preparation and enzyme preparation standing grain dimension probiotic;
(1) regulate enteron aisle fungus strain balance: beneficial bacterium enters a large amount of breedings in the enteron aisle, and antagonism is harmful to bacterium, and final field planting forms the fungus strain balance on the intestines wall, suppresses harmful bacterium and invades, and produces the material that helps to digest, promote growth in a large number simultaneously.
(2) effect of digestive ferment: all kinds of digestive ferment content that all kinds of digestive ferments and beneficial bacterium produce in metabolic activity in the preparation meet or exceed the external source interpolation level of common complex enzyme for feed.Therefore, feed is is fully digested and assimilated.
One, evidence adds 100mg/kg standing grain dimension probiotic in daily ration of broiler, can make rate of body weight gain improve 5%-9%; Add the 100mg/kg probiotic in layer diets, the egg production of laying hen can improve 5%; Be used to prevent and treat the white scour of chicken, the incidence of disease is starkly lower than 27.5%.The chick average daily gain of 30 ages in days improves 12.26%, and its protein quality of the egg that laying hen produced obviously improves, and the cholesterol level of yolk obviously reduces.
Two, the microorganism in the standing grain dimension probiotic can be synthesized nutriments such as B family vitamin, lysine, methionine, simultaneously, also the digestion that multiple digestive ferments such as protease, amylase, phytase promote feed be can produce, thereby the laying rate and the price of deed of laying hen improved.
Three, standing grain dimension probiotic can also produce the organic acid antibiotic, and kill pathogenic bacteria plays the effect of preventing and curing diseases.The enzyme of the amino acid oxidase of its generation and branch system sulfide, but harmful substances such as cracked ammonium, hydrogen sulfide, indoles reduce content of harmful gas in the hen house, reduce stink, improve environment, minimizing chicken group's the incidence of disease.Aquaculture is used standing grain dimension probiotic, not only can regulate the microecological balance of body, and promote body to nutrient absorbing, and can not cause problems such as the medicament residue and the resistance to the action of a drug, be to get a good chance of replacing antibiotic feed addictive.
Four, this feed addictive of standing grain dimension probiotic ruminant of can directly feeding, promote lumen fermentation, improve the digestion of nutritional labeling in cud, effectively improve the utilization rate of feed and the premunition of animal with synthetic again, for milk cow, can obviously improve its output of milk.This product possesses safety, characteristics of high efficiency, meets the trend of modern nutrition regulation type additive development, and after the realization industrialization is produced, expectation will be made significant contribution to the raising of China's milk cattle cultivating level.
The specific embodiment
Filamentous fungi (strain improvement and bacterial classification are cultivated)--28～32 ℃ of bacterial classification inoculations--raw material (corn, rice bran, beans, bloom)----fermentation tank--fermentation----40～50h--thalline enrichment (centrifuge)-vacuum freeze drier-low temperature drying----packing machine---finished product
------------------40～50 ℃----35～50 ℃--------40～60 orders sieve---finished product is pulverized in oven dry to Jia inserts (1: 1) to fermentation tank to raw material (corn, rice bran, bloom, beans) to bacterial classification to 28～35 ℃ of lactic acid bacterias (thalline seed selection and thalline are cultivated) in fermentation in inoculation.
Test method:
The method of sampling: every batch of sampling of product 5 casees, every case is taken out 1 bottle, and sterile working is drawn 5 milliliters for every bottle and is together put into aseptic triangular flask, shakes evenly, and adhesive label on the bottle shows name of product, Date of Sampling, lot number, sampling people, and is for inspection purpose.
The mensuration of sense index: the range estimation nasil is as good as stink, yellowish-brown liquid
The mensuration of effective viable count and assorted bacterium rate
The mensuration of bacillus subtilis viable count
The preparation of agar medium of bacillus subtilis raji cell assay Raji:
Glucose 0.5g
(NH4)SO4 0.2g
Natrium citricum 0.1g
K2HPO4 1.4g
KH2PO4 0.6g
MgSO4 0.02g
Distilled water 100ml
Purified agar 2g
pH 7.0
Confirmatory test:
Bacillus subtilis is the colony growth morphological feature on agar medium, and bacterium colony is bigger, diameter 3-10mm, and canescence, opaque, rough surface is like ground-glass appearance, and the edge often is the expansion shape.
Draw in the 45ml sterilized water that the 5ml1 product adds the band bead with aseptic straw, leave standstill 20min after, fully shake 30min, be mixed into the bacteria suspension of dilution in 1: 10, dilution obtains 1: 1 * 10 respectively so successively
2, 1: 1 * 10
3With 1: 1 * 10
4Isoconcentration (each dilution factor must be changed aseptic straw).
Draw the equal suspension 0.1ml of different dilution factors respectively with the 1ml aseptic straw, the bacillus subtilis raji cell assay Raji that adds to diameter and be the 9cm plate is used the agar medium surface, with aseptic glass slicker bacteria suspension is applied to agar surface equably.Each sample is got 3 continuously suitable dilution factors, and each dilution factor repeats 3 times, adds the blank of sterilized water simultaneously, aerobic cultivation 2-4 days, cultivation temperature are 28 ℃-30 ℃, and each dilution factor is got the thalline of 5-10 bacterium colony, smear staining, microscopic examination identification back counting bacterium colony.
The lactobacillus acidophilus viable count is measured
Lactobacillus acidophilus is cultivated on agar medium, grows up to bacterium colony, counting.The morphological feature of its bacterium colony is: for yellow, bacterium colony median size is the cotton-wool shape at the bottom of the plate, and micro white is moistening, and the edge is irregular, diameter 3 ± 1mm.Counting step is as follows:
(1) draw in the 45ml sterilized water that the 5ml2 product adds the band bead with aseptic straw, leave standstill 20min after, fully shake 30min, be mixed into the bacteria suspension of dilution in 1: 10, dilution obtains 1: 1 * 10 respectively so successively
2, 1: 1 * 10
3With 1: 1 * 10
4Isoconcentration (each dilution factor must be changed aseptic straw).
(2) drawing different dilution factor bacteria suspension 0.1ml respectively with the 1ml aseptic straw, to add to diameter be that the lactobacillus acidophilus cells of 9cm plate measures and uses the agar medium surface, with aseptic glass slicker bacteria suspension is applied to agar surface equably.Each sample is got 3 continuously suitable dilution factors, and each dilution factor repeats 3 times, adds the blank of doing of sterilized water simultaneously, anaerobism was cultivated 2～4 days, and cultivation temperature is 28 ℃～30 ℃, and each dilution factor is got the thalline of 5～10 bacterium colonies, smear staining, microscopic examination identification back counting bacterium colony.
The lactobacillus acidophilus cells measures the preparation with agar medium
Tomato juice 50ml
Yeast extract 5g
Meat extract 10g
Lactose 20g
Glucose 2g
Dipotassium hydrogen phosphate 2g
Tween 80 1g
Sodium acetate 5g
Agar 15g
Water adds to 1000ml PH 6.8 ± 0.2
Lactobacillus acidophilus is the colony growth morphological feature on agar medium, is yellow at the bottom of the plate, bacterium colony median size, and micro white, moistening, the edge is irregular, and diameter 3 ± 1mm is as cotton-wool shape bacterium colony.
The streptococcus fecalis viable count is measured
Streptococcus fecalis is cultivated on agar medium, grows up to bacterium colony, counting.The morphological feature of its bacterium colony is: bacterium colony takes on a red color or pink, and neat in edge often is ellipse or crystalline lens below the surface.Its counting step is as follows:
(1) draw in the 45ml sterilized water that the 5ml2 product adds the band bead with aseptic straw, leave standstill 20min after, fully shake 30min, be mixed into the bacteria suspension of dilution in 1: 10, dilution obtains 1: 1 * 10 respectively so successively
2, 1: 1 * 10
3With 1: 1 * 10
4Isoconcentration (each dilution factor must be changed aseptic straw).
(2) drawing the equal suspension 0.1ml of different dilution factors respectively with the 1ml aseptic straw, to add to diameter be that the streptococcus fecalis raji cell assay Raji of 9cm plate is used the agar medium surface, with aseptic glass slicker bacteria suspension is applied to agar surface equably.Each sample is got 3 continuously suitable dilution factors, and each dilution factor repeats 3 times, adds the blank of sterilized water simultaneously, anaerobism was cultivated 2～4 days, and cultivation temperature is 28 ℃～30 ℃, and each dilution factor is got the thalline of 5～10 bacterium colonies, smear staining, microscopic examination identification back counting bacterium colony.
The preparation of agar medium of streptococcus fecalis raji cell assay Raji
Peptone 10.0g
Yeast extract 10.0g
Sodium chloride 5.0g
Sodium glycero-phosphate 10.0g
Maltose 20.0g
Lactose 1.0g
Sodium azide 0.4g
Agar 20.0g
Distilled water 1000ml
Streptococcus fecalis is the colony growth morphological feature on agar medium, the red or pink bacterium colony of growing on the agar plate, and neat in edge often is ellipse or crystalline lens below the surface, all be streptococcus fecalis.
The mensuration of assorted bacterium rate
Draw in the 45ml sterilized water that the 5ml product adds the band bead with aseptic straw, leave standstill 20min after, fully shake 30min, be mixed into the bacteria suspension of dilution in 1: 10, dilution obtains 1: 1 * 10 respectively so successively
2, 1: 1 * 10
3With 1: 1 * 10
4Isoconcentration (each dilution factor must be changed aseptic straw).
Add to the agar medium surface that diameter is the 9cm plate 4.3.4.2 draw different dilution factor bacteria suspension 0.1ml respectively, bacteria suspension is applied to agar surface equably with aseptic glass slicker with the 1ml aseptic straw.Each sample is got 3 continuously suitable dilution factors, and each dilution factor repeats 3 times, adds the blank of sterilized water simultaneously, aerobic cultivation 2～4 days, and cultivation temperature is 30～33 ﹠amp; Ordm; C, each dilution factor is got the thalline of 5～10 bacterium colonies, smear staining, microscopic examination identification back counting bacterium colony.
Computing formula:
The calculating of each viable count is all carried out as follows, and statistics is calculated bacterium colony average on 3 plates of same dilution factor.
Product viable count=bacterium colony average * extension rate * 10
Assorted bacterium rate (%)=[assorted bacterium is counted ÷ (effectively bacterium number+assorted bacterium number)] * 100
Judge according to colony characteristics, calculate the specified microorganisms cell number (representing) in the product respectively with hundred million/ml.
PH value is measured
Instrument: beaker, acidometer.
Assay method:
Get the 10ml sample at random and place in the clean beaker, after preheating PH meter rectified an instrument with standard liquid (PH 6.86), the PH of working sample stirred while surveying, the stable back of instrument readings record.Each sample triplicate.
Exfactory inspection
Test stone:
Feed grade standing grain dimension probiotic is tested by the engineering department of company, should guarantee that all the feed grade standing grain that dispatches from the factory dimension probiotics all reach this standard-required, every batch of feed level microbe additive that dispatches from the factory all should be with the Certificate Of Conformance of certain format, and feed grade standing grain dimension probiotic is a collection of (3t) with the product of each fermentation tank production.
Test rating:
The exfactory inspection project that is determined as of product sensory index, effective viable count and mercury, cadmium, arsenic, lead, Escherichia coli, salmonella, mould, section tests by this standard test method by quality inspection.Be up to the standards and sign and issue the quality certification after can dispatch from the factory.
When if assay does not meet standard code, should choose sample in accordance with regulations and test to double quantity, as still not meeting standard code, this batch product is judged to defective.
Claims (5)
1. cultivate probiotics by Mixed Microbes for one kind with microorganism live bacteria preparation and enzyme preparation.
2. probiotics according to claim 1 is characterized in that what Mixed Microbes was made up of one or more bacterium of Bacillus acidi lactici bacillus, saccharomycete, filamentous fungi and photosynthetic bacteria.
3. probiotics according to claim 1 is characterized in that the culture medium of Mixed Microbes is that main material adds a small amount of assistant agent and micro-the composition by analysis for soybean powder, corn, rice bran, dregs of beans.
4. probiotics according to claim 1 is characterized in that this probiotics only contains viable bacteria or comprises thalline and the active bacteria formulation of metabolite.
5. the preparation method of probiotics as claimed in claim 1 comprises the steps:
(1) bacterial classification is selected: select 1. can assimilate matrix carbon source and inorganic nitrogen-sourced well; 2. reproduction speed is fast, mycoprotein content height; 3. avirulence and pathogenic; 4. bacterial classification stable performance; 5. preferably can mix bacterium cultivates;
(2) bacterial classification training is grown: adopt many bacterial classifications to mix bacterium and cultivate and require that strain fermentation is a raw material with the thick starch and the slag dregs of rice between bacterial strain, exist between bacterial strain and live together or laterality life or symbiosis, we adopt good bacterium by solid fermentation crude starch material and slag cypress class raw material;
(3) culture medium is that main material adds a small amount of assistant agent and trace element by analysis for soybean powder, corn, rice bran, dregs of beans;
(4) fermentation and processing bacterial classification toward seed fermentation, are inoculated into 28 ℃～32 ℃ fermentations in the fermentation tank in above-mentioned culture medium; Fermented 40-50 hour, and filtered 30-50 ℃ of oven dry of filter cake, pulverize; 40-60 sieves, photosynthetic bacteria carries out seeing under 20 ℃ of environment of light; Add minerals and vitamins, mix and make.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2005101104865A CN1965673A (en) | 2005-11-17 | 2005-11-17 | Micro-ecological preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2005101104865A CN1965673A (en) | 2005-11-17 | 2005-11-17 | Micro-ecological preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1965673A true CN1965673A (en) | 2007-05-23 |
Family
ID=38074790
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2005101104865A Pending CN1965673A (en) | 2005-11-17 | 2005-11-17 | Micro-ecological preparation |
Country Status (1)
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103652329A (en) * | 2013-12-18 | 2014-03-26 | 大连金砣水产食品有限公司 | Compound probiotic biological agent |
| CN104824367A (en) * | 2015-05-29 | 2015-08-12 | 郭心仪 | Preparation method of turtle feed |
| CN105192379A (en) * | 2015-10-08 | 2015-12-30 | 南涧县兴农发展有限公司 | Ecological-type high-quality concentrated feed for Wuliang mountain silky fowl |
| CN105255768A (en) * | 2015-11-02 | 2016-01-20 | 石河子大学 | Preparation method of lactobacillus special for milk |
| CN105360613A (en) * | 2015-11-26 | 2016-03-02 | 玉溪双胞胎饲料有限公司 | Novel green feed additive capable of replacing antibiotics |
| CN106085922A (en) * | 2016-08-12 | 2016-11-09 | 中山市润泽生物科技有限公司 | Small-sized microbial ecosystem method prepares multifunctional composite microbe microbial inoculum and application |
| CN108813223A (en) * | 2018-07-03 | 2018-11-16 | 珠海市西科水产养殖有限公司 | A kind of aquaculture specific complex microorganism formulation and preparation method thereof |
| CN108991226A (en) * | 2018-07-28 | 2018-12-14 | 陕西康大饲料有限公司 | A kind of Tiny ecosystem biological feedstuff and preparation method thereof |
-
2005
- 2005-11-17 CN CNA2005101104865A patent/CN1965673A/en active Pending
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103652329A (en) * | 2013-12-18 | 2014-03-26 | 大连金砣水产食品有限公司 | Compound probiotic biological agent |
| CN104824367A (en) * | 2015-05-29 | 2015-08-12 | 郭心仪 | Preparation method of turtle feed |
| CN105192379A (en) * | 2015-10-08 | 2015-12-30 | 南涧县兴农发展有限公司 | Ecological-type high-quality concentrated feed for Wuliang mountain silky fowl |
| CN105255768A (en) * | 2015-11-02 | 2016-01-20 | 石河子大学 | Preparation method of lactobacillus special for milk |
| CN105360613A (en) * | 2015-11-26 | 2016-03-02 | 玉溪双胞胎饲料有限公司 | Novel green feed additive capable of replacing antibiotics |
| CN106085922A (en) * | 2016-08-12 | 2016-11-09 | 中山市润泽生物科技有限公司 | Small-sized microbial ecosystem method prepares multifunctional composite microbe microbial inoculum and application |
| CN108813223A (en) * | 2018-07-03 | 2018-11-16 | 珠海市西科水产养殖有限公司 | A kind of aquaculture specific complex microorganism formulation and preparation method thereof |
| CN108991226A (en) * | 2018-07-28 | 2018-12-14 | 陕西康大饲料有限公司 | A kind of Tiny ecosystem biological feedstuff and preparation method thereof |
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Open date: 20070523 |