CN106190933A - The bacillus subtilis of the anti-pathogenic bacterium of wide spectrum and application thereof - Google Patents
The bacillus subtilis of the anti-pathogenic bacterium of wide spectrum and application thereof Download PDFInfo
- Publication number
- CN106190933A CN106190933A CN201610857510.XA CN201610857510A CN106190933A CN 106190933 A CN106190933 A CN 106190933A CN 201610857510 A CN201610857510 A CN 201610857510A CN 106190933 A CN106190933 A CN 106190933A
- Authority
- CN
- China
- Prior art keywords
- bacillus subtilis
- reagent
- application
- jlth
- terms
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
Abstract
The present invention relates to bacillus subtilis and the application thereof of the anti-pathogenic bacterium of a kind of wide spectrum, belong to microbial technology field.A bacillus subtilis disclosed by the invention, its deposit number is CGMCC 11164.There is the ability of wide spectrum suppression pathogenic bacterium (predominantly escherichia coli, staphylococcus aureus, Salmonella typhimurium, shigella, α-hemolytic streptococcus, beta hemolytic streptococcus), and there is the strongest product protease, amylase and the ability of cellulase, this bacterium has good application prospect in terms of feedstuff and Tiny ecosystem reagent.
Description
Technical field
The present invention relates to bacillus subtilis and the application thereof of the anti-pathogenic bacterium of a kind of wide spectrum.
Background technology
In recent years, limit along with many is national in the world or forbids using antibiotic in feedstuff.Find new antibiotic
Succedaneum becomes a urgent task of animal husbandry.People start to seek one after another other succedaneum and substitute technology, to protect
The efficiency of card Animal husbandry production is unaffected with benefit.Fermented feed and microbial ecological agent technology are having of recently growing up
The New production technology of many advantages, feedstuff contains more active probiotic after probiotics fermention and interpolation microbial ecological agent
Thalline, various enzyme, metabolite at different levels, multivitamin, protein breakdown products, active small peptide, aminoacid, antibacterial substance,
The immunostimulant factor, somatomedin etc., play the colony balance effect promoting growth, maintaining animal intestinal；Due to without
Antibiotic etc, does not results in antibiotics residues, adds probiotics micro-ecological formulation and fermented feed is a kind of ecological
Health type Feed Production Technology.
Bacillus subtilis is the feed microbe strain that China allows to use, and because it is nontoxic, harmless, is often used in and changes
Kind animal intestinal function, promotion growth of animal and prevention disease.Develop novel Bacillus subtilis strain, beneficially China's herding
Industry develops in a healthy way, it is ensured that people's food safety.
Summary of the invention
It is an object of the invention to provide bacillus subtilis and the application thereof of the anti-pathogenic bacterium of a kind of wide spectrum, described hay bud
Spore bacillus (Bacillus subtilis) JLTH-146 screening, from the fresh excrement sample of sodium selenite, has wide spectrum suppression pathogenic bacterium
Ability, and have and the strongest produce protease, amylase and the ability of cellulase, can be as fermented feed leaven and micro-life
State formulation application is in livestock and poultry cultivation, to make up the deficiencies in the prior art.
The above-mentioned purpose of the present invention is achieved through the following technical solutions:
The bacillus subtilis of the anti-pathogenic bacterium of wide spectrum, described bacillus subtilis is preserved in Chinese micro-life on July 31st, 2015
Thing culture presevation administration committee's common micro-organisms center, preserving number is CGMCC 11164.
Described bacillus subtilis in terms of feedstuff in application.
Described bacillus subtilis is applied to fermented feed leaven field.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application, the preparation of described Tiny ecosystem reagent is:
(1), prepared by seed liquor: chooses the bacillus subtilis JLTH-146 bacterium mud that a ring is fresh, is inoculated in 100mLLB culture medium
In, 500mL triangle bottled liquid 200mL, constant temperature 37 DEG C, 200rpm cultivate 12-16h seed liquor；
(2), work fermentation liquid is cultivated: above-mentioned seed liquor be seeded in sterilized liquid LB culture medium with 8-12% inoculum concentration, constant temperature
37 DEG C, 200r/min cultivate 16-24h, plate count is 108~1010Fermentation culture is stopped during CFU/mL；
(3), prepared by mycopowder: is centrifuged by work fermentation liquid, 10000rpm, 8min, collects thalline, adds dextrin stirring and evenly mixing, warp
Cross vacuum lyophilization and become powder, make containing the bacillus subtilis microbial ecological agent that viable bacteria amount is 100-1000 hundred million/g.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application, described microorganism reagent is bacillus subtilis
The mycopowder of bacterium JLTH-146, is that the ratio of 0.1% ~ 1.0% is added to animal and fowl fodder by mass percentage.
The beneficial effects of the present invention is: bacillus subtilis (Bacillus subtilis) JLTH-146 of the present invention
By 16S rDNA sequence alignment analysis, with the bacillus subtilis Bacillus of GenBank accession number NR 102783.1
The homology of the 16S rDNA sequence of subtilis strain 168 bacterial strain is the highest, and similarity is 99%, by combining thalline shape
State feature, growth conditions, Physiology and biochemistry qualification result determine that bacillus subtilis JLTH-146 belongs to bacillus
(Bacillus), for bacillus subtilis (Bacillus subtilis).
Bacillus subtilis JLTH-146 of the present invention has broad-spectrum antibacterial action, to various pathogens (predominantly large intestine bar
Bacterium, staphylococcus aureus, shigella, Salmonella typhimurium, α-hemolytic streptococcus, beta hemolytic streptococcus)
There is good inhibiting effect.
The bacillus subtilis JLTH-146 of the present invention is applied to fermented feed leaven field.Described fermented feed ferments
Agent is bacillus subtilis JLTH-146 liquid fermentation liquid, adds fermentation materials to by feeding quality percentage ratio 1%-5% to be fermented
In.
Another aspect of the present invention provides a kind of microbial ecological agent comprising above-mentioned bacillus subtilis JLTH-146.Institute
The microbial ecological agent stated is the mycopowder of bacillus subtilis JLTH-146, is that the ratio of 0.1%-1.0% is added by mass percentage
To animal and fowl fodder.
The bacillus subtilis JLTH-146 of the present invention has wide spectrum suppression pathogenic bacterium ability and the highest enzymatic productivity, can
It is applied in animal and fowl fodder as feed fermentation agent and Tiny ecosystem reagent, the production performance of cultivated animals can be effectively improved, application
Have a extensive future.
Accompanying drawing explanation
Accompanying drawing described herein is used for providing a further understanding of the present invention, constitutes the part of the application, this
Bright illustrative example and explanation thereof are used for explaining the present invention, are not intended that inappropriate limitation of the present invention.
Fig. 1 is the bacillus subtilis JLTH-146 gene order of the present invention；
Fig. 2 is the bacillus subtilis JLTH-146 phylogenetic tree of the present invention.
Detailed description of the invention
Further illustrate detailed content and the detailed description of the invention thereof of the present invention below in conjunction with the accompanying drawings.
Seeing shown in Fig. 1 and Fig. 2, bacillus subtilis described in the bacillus subtilis of the anti-pathogenic bacterium of wide spectrum of the present invention exists
On July 31st, 2015 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC
11164。
Described bacillus subtilis in terms of feedstuff in application.
Described bacillus subtilis is applied to fermented feed leaven field.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application, the preparation of described Tiny ecosystem reagent is:
(1), prepared by seed liquor: chooses the bacillus subtilis JLTH-146 bacterium mud that a ring is fresh, is inoculated in 100mLLB culture medium
In, 500mL triangle bottled liquid 200mL, constant temperature 37 DEG C, 200rpm cultivate 12-16h seed liquor；
(2), work fermentation liquid is cultivated: above-mentioned seed liquor be seeded in sterilized liquid LB culture medium with 8-12% inoculum concentration, constant temperature
37 DEG C, 200r/min cultivate 16-24h, plate count is 108~1010Fermentation culture is stopped during CFU/mL；
(3), prepared by mycopowder: is centrifuged by work fermentation liquid, 10000rpm, 8min, collects thalline, adds dextrin stirring and evenly mixing, warp
Cross vacuum lyophilization and become powder, make containing the bacillus subtilis microbial ecological agent that viable bacteria amount is 100-1000 hundred million/g.
Described bacillus subtilis in terms of Tiny ecosystem reagent in application, described microorganism reagent is bacillus subtilis
The mycopowder of bacterium JLTH-146, is that the ratio of 0.1% ~ 1.0% is added to animal and fowl fodder by mass percentage.
Embodiment:
Experimental technique used in following embodiment if no special instructions, is conventional method.Used by following embodiment
Material, reagent etc., if no special instructions, the most commercially obtain.
LB culture medium: peptone 10g, yeast powder 3g, sodium chloride 5g, glucose 3g, water 1000mL.pH6.2-7.0.Nutrition
Agar plate: peptone 10g, Carnis Bovis seu Bubali cream 3g, sodium chloride 5g, agar 15g, distilled water 1000mL.PH to 7.2～7.4.
Blood plate culture medium: peptone 10g, Carnis Bovis seu Bubali cream 3g, de-fiber Sanguis caprae seu ovis 50mL, sodium chloride 5g, agar 15g, steaming
Distilled water 1000mL.PH to 7.2～7.4.
Embodiment 1
Bacillus subtilis separation screening and qualification:
1, bacterial strain primary dcreening operation
Gather 20 parts of various places, Jilin Province sodium selenite fresh excrement sample, weigh 5g sample respectively, add 45mL physiological saline solution, system
Standby one-tenth sample suspension, after 80 DEG C of water bath processing, coats on LB nutrition flat board, and constant temperature 37 DEG C is cultivated 48 hours, it is thus achieved that 168 strains
Bacillus cereus.
2, suppression pathogenic bacterium bacillus cereus Preliminary screening
By the bacterial strain of isolated, after constant temperature 37 DEG C is cultivated 16-48 hour, 8000r/min is centrifuged 10min, takes supernatant, will filter
Scraps of paper diameter 0.8cm is soaked in 15min in the supernatant of isolated, take escherichia coli (Escherichiacoli)
CMCC44825 is indicator bacteria 50 μ L, is spread evenly across on nutrient agar panel, stands 20min, puts ready filter paper,
Constant temperature 37 DEG C is cultivated 16 hours, observes antibacterial situation.Survey the diameter of inhibition zone with slide gauge, represent with the diameter of inhibition zone and press down
Bacterium activity.Escherichia coli are purchased from Guangdong microorganism germ plasma resources bank.
Through bacteriostatic experiment, screening obtains the bacterial strain that 1 strain is the highest to escherichia coli inhibition, identifies this bacterial strain further
Analyze.
3, bacillus cereus taxonomy is identified
(1) according to " the outstanding Bacteria Identification handbook of uncle " (the 8th edition) and " common bacteria system identification handbook " (the elegant pearl in east, Cai Miaoying
Etc. writing, Beijing: Science Press, 2001.2) method that described in, bacterium classification is identified, carries out form to bacterial strain JLTH-146
Feature, cultural character and physio-biochemical characteristics are identified, concrete outcome is as follows:
The form of thalline and physio-biochemical characteristics:
When sporeformer grows on LB flat board, bacterium colony is relatively big, white micro-Huang, circular, and center projections is translucent, cultivates for a long time
Easily form fold on surface, Grain-positive, thalline is rod-short (0.5～0.6) μ m (1.3～1.6) μm, and spore is ellipse
Life in circle.
Physiological and biochemical property: result is as shown in table 1.
Table 1 bacillus subtilis JLTH-146 physiological and biochemical test
| Detection project | Result |
| Gram’s staining | + |
| To oxygen demand | + |
| Casein hydrolysis | + |
| Starch Hydrolysis | + |
| Gelatin liquefaction | + |
| Nitrate reduction | + |
| Citrate test | + |
| Catalase test | + |
| Litmus milk reduces | + |
| V P measures | + |
| Carbamide utilizes | - |
| Produce indole test | - |
| Glucose | + |
| Fructose | + |
| Lactose | + |
| Xylose | + |
| Mannose | + |
| 9%NaCl | + |
Remarks :-represent negative reaction；+ represent positive reaction.
(2) 16S rDNA test
Extract the STb gene of JLTH-146, with it as template, utilize bacterial 16 S rDNA universal primer to carry out PCR amplification, grown
Degree is about the amplified production of 1.5kb, is reclaimed by amplified production, entrusts Shanghai Sangon Biological Engineering Technology And Service Co., Ltd's order-checking
Company checks order.The sequence recorded is as shown in Figure 1.
Sequencing result is compared with Gen-Bank sequence homology, then with software MEGA5.0 phylogenetic tree construction (as
Shown in Fig. 2), to determine the race relation of bacterial strain.Homology analysis result shows, bacterial strain JLTH-146 and Bacillus
Subtilis strain 168 (GenBank accession number NR 102783.1) homology is 99%, and it is same to be in phylogenetic tree
One branch.
Based on morphological features, growth conditions, Physiology and biochemistry, 16S rDNA qualification result feature, by bacterial strain JLTH-
146 are accredited as Bacillus subtilis.The Classification And Nomenclature of suggestion is bacillus subtilis；This bacterial strain is July 31 in 2015
Day it is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center and (is called for short CGMCC 9501, address: Beijing
North Star West Road, Chaoyang District 1 institute 3, Institute of Microorganism, Academia Sinica, postcode 100101), preserving number is CGMCC
No.11164。
Embodiment 2: bacillus subtilis JLTH-146 suppression pathogenic bacterium experiment
1, suppression pathogenic bacterium ability detection:
By the bacterial strain of isolated, after constant temperature 37 DEG C is cultivated 16-48 hour, 8000r/min is centrifuged 10min, takes supernatant, will filter
Scraps of paper diameter 0.8cm is soaked in 15min in the supernatant of isolated, take escherichia coli (Escherichiacoli)
CMCC44825, staphylococcus aureus (Staphylococcusaureus) CMCC26071, Salmonella typhimurium
(Salmonellatyphimurium) CMCC50, shigella (Shigellaflexneri) CMCC51061 is that indicator bacteria respectively takes
50 μ L, are spread evenly across on nutrient agar panel.α-hemolytic streptococcus (α-hemolytic streptococcus)
CMCCB32213, beta hemolytic streptococcus (β-hemolytic streptococcus) CMCCB32210 is that indicator bacteria respectively takes
50 μ L, are spread evenly across in blood plate culture medium.Standing 20min, put ready filter paper, it is little that constant temperature 37 DEG C cultivates 16
Time, observe antibacterial situation.Survey the diameter of inhibition zone with slide gauge, represent bacteriostatic activity with the diameter of inhibition zone.Escherichia coli,
Staphylococcus aureus, Salmonella typhimurium, shigella are purchased from Guangdong microorganism germ plasma resources bank.A type hemolytic hammer
Bacterium, beta hemolytic streptococcus are purchased from Chinese medicine antibacterial preservation administrative center.
Bacillus subtilis JLTH-146 suppression pathogenic bacterium ability is as shown in table 2:
Table 2 hay spore pipe bacterium JLTH-146 suppresses pathogenic bacterium ability test
Embodiment 3: bacillus subtilis JLTH-146 produces enzyme experiment
1, α-amylase detection:
Starch culture-medium: peptone 10g, Carnis Bovis seu Bubali cream 5g, NaCl 5g, soluble starch 2g, distilled water 1000ml, agar powder
20g。
By in the bacterial strain dibbling of isolated to culture medium, after 37 DEG C are cultivated 16-48 hour, in the training that strain growth is good
Support primary surface and drip a certain amount of iodine liquid so that it is be uniformly covered with whole culture medium flat plate, if nothing occurs in the bacterium colony periphery grown
Color transparent circle, illustrates that starch is hydrolyzed.Measure the ratio of transparent circle diameter and colony diameter.
2, protease detection:
Milk culturemedium: skim milk 50ml, distilled water 50ml, agar powder 1.5g.
By the bacterial strain dibbling arrived of separation to milk culturemedium, cultivate 16-48 hour for 37 DEG C, if the bacterium colony week grown
While there is transparent circle to produce, illustrate that in culture medium, casein has been hydrolyzed.Measure the ratio of transparent circle diameter and colony diameter.
3, cellulase detection:
Cellulose culture medium: Nutrient agar+0.5%CMC:
By the bacterial strain dibbling of isolated to cellulose culture medium, cultivate 16-48 hour, make it with 0.1% Congo red solution for 37 DEG C
Uniformly it is covered with whole culture medium flat plate, dyes 30 minutes, then drip a certain amount of 1moL/L sodium chloride, be covered with culture medium flat plate table
Face, decolours about 20 minutes, if hydrolysis occurs in the bacterium colony periphery grown, illustrates that in culture medium, CMC is by water
Solve.
Measure the ratio of transparent circle diameter and colony diameter.
The product enzyme situation of bacillus subtilis JLTH-146 is as shown in table 3:
The product enzyme test of table 3 bacillus subtilis JLTH-146
By in table it can be seen that bacillus subtilis JLTH-146 has the strongest product amylase, protease and cellulase energy
Power, the transparent circle diameter of three kinds of enzymes and colony diameter ratio all about 5.0, therefore the present invention screening bacillus subtilis
JLTH-146 has the biggest potentiality in the application of fermented feed and animal microecological formulation thereof.
Embodiment 4: the preparation of bacillus subtilis JLTH-146 fermented feed
1, prepared by seed liquor: chooses the bacillus subtilis JLTH-146 bacterium mud that a ring is fresh, is inoculated in 100mLLB culture medium
(250mL triangle bottled liquid 100mL), constant temperature 37 DEG C, 200r/min cultivate 16-24h seed liquor；
2, work fermentation liquid is cultivated: above-mentioned seed liquor be seeded in sterilizing LB fluid medium with 8-12% inoculum concentration, constant temperature 37
DEG C, 200r/min cultivate 16-24h, plate count is 108~1010Fermentation culture is stopped during CFU/mL；
3, prepared by fermented feed: accessed by fermentation materials quality 5% ~ 12% by the work fermentation liquid prepared, constant temperature 37 DEG C cultivation
35-48 hour, pack as early as possible after fermentation, it is desirable to material is compacted, airtight.
After normal fermentation material formula: Semen Maydis powder 62%, bean cake 22%, Testa Tritici 12%, premix material 4%, nutrient content and fermentation
Nutrient content is shown in Table 4:
Nutritional labeling change table before and after table 4 conventional pig feed fermentation
| Conventional nutrients | Before fermentation | After fermentation | Rate of change % |
| Crude protein % | 18.2% | 19.3% | +6.04% |
| Crude fibre % | 5.7% | 4.1% | -28.07% |
| Lysine | 0.82% | 0.89% | +8.53% |
As seen from the above table, the nutritive value generation significant change of conventional feed after fermentation.In feedstuff, crude fibre have dropped
28.07%, crude protein improves 6.04% on the basis of original, lysine improve 8.53% on the basis of original.Improve feedstuff
Feedstuff is digested and assimilated by nutritive value, beneficially animal.
Embodiment 5: the preparation of bacillus subtilis JLTH-146 microbial ecological agent
1, prepared by seed liquor: chooses the bacillus subtilis JLTH-146 bacterium mud that a ring is fresh, is inoculated in 100mLLB culture medium
(500mL triangle bottled liquid 200mL), constant temperature 37 DEG C, 200rpm cultivate 12-16h seed liquor；
2, work fermentation liquid is cultivated: above-mentioned seed liquor be seeded in sterilized liquid LB culture medium with 8-12% inoculum concentration, constant temperature 37
DEG C, 200r/min cultivate 16-24h, plate count is 108~1010Fermentation culture is stopped during CFU/mL；
3, prepared by mycopowder: work fermentation liquid centrifugal (10000rpm, 8min) is collected thalline, adds appropriate dextrin stirring mixed
Even, become powder through vacuum lyophilization, make containing the bacillus subtilis microbial ecological agent that viable bacteria amount is 100-1000 hundred million/g.
Embodiment 6: the bacillus subtilis JLTH-146 microbial ecological agent impact on weaned piglets
60 well-growns, piglet that body weight is close being randomly divided into 2 groups, often 3 repetitions of group, each repetition 10, I group is right
According to group, feeding general goods daily ration, II group is experimental group, and adding mass percent on the basis of general goods daily ration is 0.2%
Bacillus subtilis JLTH-146 microbial ecological agent.Experimental period is 40 days, measures the average daily gain of piglet, feedstuff-meat ratio and diarrhoea
Rate.Experimental result is as shown in table 5:
The impact on weaned piglets of the table 5 bacillus subtilis JLTH-146 microbial ecological agent
| Group | Average daily gain (g) | Feedstuff-meat ratio F/G | Diarrhea rate (%) |
| Ⅰ | 311.11±14.22 | 1.99±0.01 | 17.52±1.71 |
| Ⅱ | 339.27±12.17 | 1.74±0.04 | 3.74±1.56 |
By result of the test in upper table it can be seen that feeding trial meat on the 40th, II group of (experimental group) piglet average daily gain ratio I group
(matched group) increases 28.16g, and piglet feedstuff-meat ratio reduces by 0.25, and diarrhea rate reduces by 13.7%.This result of the test shows to add in daily ration
Add bacillus subtilis and can significantly improve the production performance of piglet cultivation, reduce daily nursing cost, increase raiser's economy and receive
Benefit.Bacillus subtilis is added with good application prospect in pig starter feed.
The foregoing is only the preferred embodiment of the present invention, be not limited to the present invention, for the technology of this area
For personnel, the present invention can have various modifications and variations.All any modification, equivalent substitution and improvement made for the present invention etc.,
Should be included within the scope of the present invention.
Claims (6)
1. the bacillus subtilis of the anti-pathogenic bacterium of wide spectrum, it is characterised in that: described bacillus subtilis was July 31 in 2015
Day is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preserving number is CGMCC 11164.
Bacillus subtilis the most according to claim 1 in terms of feedstuff in application.
Bacillus subtilis the most according to claim 1 is applied to fermented feed leaven field.
Bacillus subtilis the most according to claim 1 in terms of Tiny ecosystem reagent in application.
Bacillus subtilis the most according to claim 1 in terms of Tiny ecosystem reagent in application, described Tiny ecosystem reagent
Preparation be:
(1), prepared by seed liquor: chooses the bacillus subtilis JLTH-146 bacterium mud that a ring is fresh, is inoculated in 100mLLB culture medium
In, 500mL triangle bottled liquid 200mL, constant temperature 37 DEG C, 200rpm cultivate 12-16h seed liquor；
(2), work fermentation liquid is cultivated: above-mentioned seed liquor be seeded in sterilized liquid LB culture medium with 8-12% inoculum concentration, constant temperature
37 DEG C, 200r/min cultivate 16-24h, plate count is 108~1010Fermentation culture is stopped during CFU/mL；
(3), prepared by mycopowder: is centrifuged by work fermentation liquid, 10000rpm, 8min, collects thalline, adds dextrin stirring and evenly mixing, warp
Cross vacuum lyophilization and become powder, make containing the bacillus subtilis microbial ecological agent that viable bacteria amount is 100-1000 hundred million/g.
Bacillus subtilis the most according to claim 1 in terms of Tiny ecosystem reagent in application, described microorganism reagent
For the mycopowder of bacillus subtilis JLTH-146, it is that the ratio of 0.1% ~ 1.0% is added to animal and fowl fodder by mass percentage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610857510.XA CN106190933B (en) | 2016-09-28 | 2016-09-28 | The bacillus subtilis of the anti-pathogenic bacteria of wide spectrum and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610857510.XA CN106190933B (en) | 2016-09-28 | 2016-09-28 | The bacillus subtilis of the anti-pathogenic bacteria of wide spectrum and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106190933A true CN106190933A (en) | 2016-12-07 |
| CN106190933B CN106190933B (en) | 2019-08-13 |
Family
ID=57520977
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610857510.XA Active CN106190933B (en) | 2016-09-28 | 2016-09-28 | The bacillus subtilis of the anti-pathogenic bacteria of wide spectrum and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106190933B (en) |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058181A (en) * | 2017-04-13 | 2017-08-18 | 中国农业科学院特产研究所 | A kind of bacillus subtilis and its separation method and application |
| CN108660100A (en) * | 2018-07-11 | 2018-10-16 | 金福赛（北京）生物科技有限公司 | Bacillus subtilis and its application in microbial inoculum and broiler fodder |
| CN108865928A (en) * | 2018-06-13 | 2018-11-23 | 铜仁职业技术学院 | A kind of bacillus subtilis and the method for being used to prepare fruit of negundo probiotics |
| CN110272854A (en) * | 2019-07-29 | 2019-09-24 | 北京林业大学 | A kind of bacillus subtilis strain and its application |
| CN111187730A (en) * | 2019-11-28 | 2020-05-22 | 中国农业科学院兰州兽医研究所 | Bacillus subtilis and application thereof |
| CN112501062A (en) * | 2020-11-27 | 2021-03-16 | 吉林省农业科学院 | Bacillus subtilis for producing levansucrase and application thereof |
| CN113462597A (en) * | 2021-06-29 | 2021-10-01 | 广西壮族自治区兽医研究所 | Bacillus subtilis for producing lignin peroxidase, microbial inoculum containing bacillus subtilis and application of bacillus subtilis |
| CN113913325A (en) * | 2021-09-10 | 2022-01-11 | 河北省畜牧兽医研究所 | Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application thereof |
| CN115216431A (en) * | 2022-08-09 | 2022-10-21 | 龙岩学院 | Multifunctional bacillus subtilis from corncobs and application thereof |
| CN116024114A (en) * | 2022-07-07 | 2023-04-28 | 广东容大生物股份有限公司 | Intestinal bacillus subtilis strain and preparation method and application thereof |
| CN116024146A (en) * | 2023-03-28 | 2023-04-28 | 中国农业大学 | Bacillus subtilis with high antibacterial activity and fermentation condition optimization thereof |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329749A (en) * | 2011-09-19 | 2012-01-25 | 山东宝来利来生物工程股份有限公司 | Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis |
| CN103060222A (en) * | 2012-09-19 | 2013-04-24 | 中国农业科学院饲料研究所 | Bacillus subtilis B27 with probiotic effect and application thereof |
| CN103614327A (en) * | 2013-11-27 | 2014-03-05 | 北京昕大洋科技发展有限公司 | Bacillus subtilis and use thereof |
| CN103981118A (en) * | 2013-12-24 | 2014-08-13 | 北京伟嘉人生物技术有限公司 | Bacillus subtilis feed additive and preparation method and application thereof |
| CN104109643A (en) * | 2014-04-23 | 2014-10-22 | 中国海洋大学 | Bacillus spp with broad-spectrum antibacterial activity |
| CN104774781A (en) * | 2014-08-26 | 2015-07-15 | 汕头大学 | Bacillus subtilis DCU and use thereof |
-
2016
- 2016-09-28 CN CN201610857510.XA patent/CN106190933B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102329749A (en) * | 2011-09-19 | 2012-01-25 | 山东宝来利来生物工程股份有限公司 | Bacillus subtilis bred by space mutation breeding technology and application of Bacillus subtilis |
| CN103060222A (en) * | 2012-09-19 | 2013-04-24 | 中国农业科学院饲料研究所 | Bacillus subtilis B27 with probiotic effect and application thereof |
| CN103614327A (en) * | 2013-11-27 | 2014-03-05 | 北京昕大洋科技发展有限公司 | Bacillus subtilis and use thereof |
| CN103981118A (en) * | 2013-12-24 | 2014-08-13 | 北京伟嘉人生物技术有限公司 | Bacillus subtilis feed additive and preparation method and application thereof |
| CN104109643A (en) * | 2014-04-23 | 2014-10-22 | 中国海洋大学 | Bacillus spp with broad-spectrum antibacterial activity |
| CN104774781A (en) * | 2014-08-26 | 2015-07-15 | 汕头大学 | Bacillus subtilis DCU and use thereof |
Cited By (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058181A (en) * | 2017-04-13 | 2017-08-18 | 中国农业科学院特产研究所 | A kind of bacillus subtilis and its separation method and application |
| CN107058181B (en) * | 2017-04-13 | 2020-07-28 | 中国农业科学院特产研究所 | Bacillus subtilis and separation method and application thereof |
| CN108865928A (en) * | 2018-06-13 | 2018-11-23 | 铜仁职业技术学院 | A kind of bacillus subtilis and the method for being used to prepare fruit of negundo probiotics |
| CN108865928B (en) * | 2018-06-13 | 2022-03-22 | 铜仁职业技术学院 | Bacillus subtilis and method for preparing negundo chastetree fruit microecological preparation |
| CN108660100B (en) * | 2018-07-11 | 2021-12-21 | 金福赛（北京）生物科技有限公司 | Bacillus subtilis and application thereof in microbial inoculum and broiler feed |
| CN108660100A (en) * | 2018-07-11 | 2018-10-16 | 金福赛（北京）生物科技有限公司 | Bacillus subtilis and its application in microbial inoculum and broiler fodder |
| CN110272854A (en) * | 2019-07-29 | 2019-09-24 | 北京林业大学 | A kind of bacillus subtilis strain and its application |
| CN110272854B (en) * | 2019-07-29 | 2021-07-27 | 北京林业大学 | Bacillus subtilis strain and application thereof |
| CN111187730A (en) * | 2019-11-28 | 2020-05-22 | 中国农业科学院兰州兽医研究所 | Bacillus subtilis and application thereof |
| CN113684157A (en) * | 2020-11-27 | 2021-11-23 | 吉林省农业科学院 | Bacillus subtilis for producing levansucrase and application thereof |
| CN112501062A (en) * | 2020-11-27 | 2021-03-16 | 吉林省农业科学院 | Bacillus subtilis for producing levansucrase and application thereof |
| CN113462597A (en) * | 2021-06-29 | 2021-10-01 | 广西壮族自治区兽医研究所 | Bacillus subtilis for producing lignin peroxidase, microbial inoculum containing bacillus subtilis and application of bacillus subtilis |
| CN113913325A (en) * | 2021-09-10 | 2022-01-11 | 河北省畜牧兽医研究所 | Bacillus subtilis, Ericin1a antibacterial peptide secreted by bacillus subtilis and application thereof |
| CN116024114A (en) * | 2022-07-07 | 2023-04-28 | 广东容大生物股份有限公司 | Intestinal bacillus subtilis strain and preparation method and application thereof |
| CN115216431A (en) * | 2022-08-09 | 2022-10-21 | 龙岩学院 | Multifunctional bacillus subtilis from corncobs and application thereof |
| CN115216431B (en) * | 2022-08-09 | 2023-12-12 | 龙岩学院 | Multifunctional bacillus subtilis from corncob and application thereof |
| CN116024146A (en) * | 2023-03-28 | 2023-04-28 | 中国农业大学 | Bacillus subtilis with high antibacterial activity and fermentation condition optimization thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106190933B (en) | 2019-08-13 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106190933B (en) | The bacillus subtilis of the anti-pathogenic bacteria of wide spectrum and its application | |
| CN106399196B (en) | One lactobacillus plantarum HEW-A490 and its application | |
| CN101690545B (en) | Method for producing complex micro-ecological preparation with microbial agents and enzyme | |
| JP5872104B2 (en) | New Bacillus subtilis {NOVELBACILLUSSUBTILIS} | |
| CN101715879B (en) | Compound beta-mannase and lactobacillus feed additive | |
| CN108208316B (en) | Siamese bacillus-containing feed | |
| CN101671638B (en) | New strain of bifidobacterium and fermentative preparation method and application thereof | |
| CN107164269A (en) | A kind of lactobacillus paracasei, preparation and its application in pig feed | |
| CN102640885B (en) | Applications of Lactobacillus plantarum in complete-strain Triticumspp silage | |
| CN103087964A (en) | Bacillus subtilis, microecological preparation and application of bacillus subtilis in animal feed | |
| CN104651268A (en) | Lactobacillus plantarum and application thereof | |
| CN103966118A (en) | Bacillusamyloliquefaciens and application thereof | |
| CN103070285A (en) | Microbe feed additive and preparation method thereof | |
| CN102168051B (en) | Bacillus cereus with higher resistance to jinggangmycin aqua and application | |
| CN102229902B (en) | Bacillus pumilus, feed additive thereof, premix, and feed mildewproof agent | |
| CN103525718B (en) | Bacillus cereus and probiotics powder thereof as well as preparation and application of probiotics powder | |
| CN108541805A (en) | The reuse method and fermentate of a kind of clostridium butyricum fermentation waste water and its application | |
| CN107974421A (en) | A kind of lactobacillus acidophilus and its screening technique and application, a kind of microbial inoculum | |
| CN102911901B (en) | Bacillus pumilus strain and application thereof | |
| CN102517227B (en) | Enterococcus faecalis and applications and feed additive and leavening agent thereof | |
| CN102628023B (en) | Bacillus subtilis.XD08B having high ganoderma lucidum polysaccharide yield, feeding microbial preparation prepared from bacillus subtilis.XD08B and preparation method of the feeding microbial preparation | |
| CN103266074B (en) | B.subtilis spores strain and application thereof | |
| CN102907585A (en) | Probiotics and vitamin premix and mixture for middle pig | |
| CN102919557A (en) | Growing fattening pig probiotics composite premix and accessory material | |
| CN102660534A (en) | Bacillus subtilis preparation as well as preparation method and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |