CN102965316B - Bacteriocin containing class and the feed addictive of Pediococcus acidilactici, premix and batch - Google Patents

Bacteriocin containing class and the feed addictive of Pediococcus acidilactici, premix and batch Download PDF

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CN102965316B
CN102965316B CN201210476707.0A CN201210476707A CN102965316B CN 102965316 B CN102965316 B CN 102965316B CN 201210476707 A CN201210476707 A CN 201210476707A CN 102965316 B CN102965316 B CN 102965316B
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pediococcus acidilactici
feed addictive
freeze
bacteriocin
culture
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CN102965316A (en
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胡婷
贠桂玲
李曼
韩科厅
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Wuhan agricultural farming and animal husbandry development Co., Ltd.
Beijing Dabeinong Technology Group Co Ltd
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Wuhan Agricultural Farming And Animal Husbandry Development Co Ltd
Beijing Dabeinong Technology Group Co Ltd
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Abstract

The invention discloses a kind of Pediococcus acidilactici(Pediococcus acidilactici)New strains HT, and the feed addictive containing bacillus category bacteriocin and Pediococcus acidilactici.The Pediococcus acidilactici new strains HT of the present invention, its preserving number is CGMCC No.6500.Bacillus subtilis CGMCC No.6511 class bacteriocin freeze-dried powder and Pediococcus acidilactici HT bacterium powders are proportionally sufficiently mixed, obtain feed addictive.The feed addictive is a kind of green feed additive, is improved food conversion ratio, and improves breeding performonce fo animals and immunity, prevents and treats diarrhoea, plays the effect for improving animal products quality.

Description

Bacteriocin containing class and the feed addictive of Pediococcus acidilactici, premix and batch
Technical field
Present invention relates particularly to a kind of bacteriocin containing class and the feed addictive of Pediococcus acidilactici, premix and batch, Belong to additive for microbe feedstuff field.
Background technology
Antibiotic not only solve growth it is low with premunition soon between contradiction, and bring to aquaculture huge Economic benefit so that antibiotic generally uses as feed addictive.However, long-term use of the antibiotic in feed is brought Many negative effects such as medicament residue, resistance and environmental pollution etc., endanger human health.
As food-safety problem obtains more and more extensive concern in the world, reduce or stop in husbandry sector Using the cry more and more higher of antibiotic, thus use is similar but nontoxic to antibiotic function, have no side effect, noresidue, nonreactive The property of medicine, while raw materials of feed additives also free from environmental pollution turns into a kind of inexorable trend.
In recent years, the appearance of probiotics, enzyme preparation, Chinese herbal and crude drugs preparations and organic acid additive, for solving this One problem plays a significantly greater role, but the use of many products is unsatisfactory, such as:Benefit can be played using additive for microbe feedstuff The effect of strain is filled or be inoculated with, but can not fully play a part of transferring other probiotics in enteron aisle;The benefits such as small peptide are used alone Raw first additive effect is relatively simple, can activate endogenic probiotics and play a role, but can lack with unique nutrition Function, stimulate immune and anti-disease ability special strain.In a word, perfect compound feed and can not thoroughly break away to antibiosis at present The dependence of plain quasi-growth promoter.
The content of the invention
To solve the above problems, present invention firstly provides a kind of Pediococcus acidilactici (Pediococcus acidilactici) New strains and its lyophilized bacterium powder.
The present invention also provides a kind of bacillus subtilis (Bacillus subtilis) class bacteriocin freeze-dried powder.
The present invention also provides the feed addictive containing bacillus subtilis mushroom bacteriocin and Pediococcus acidilactici, premix and Batch.
Pediococcus acidilactici (Pediococcus acidilactici) new strains HT provided by the invention is applicant from duck It is isolated in jejunal content.Biological property is as follows:Cell spherical shape, tetrad is formed in the plane alternating division of two, right angle Shape, general cell are given birth in pairs, and single survivor is rare, not into catenation.Gram-positive, do not move, amphimicrobian.Trained in MRS It is small to support bacterium colony on base, it is white.Using glucose, ribose, xylose, galactolipin, lactose, sucrose, trehalose, fructose, seven leaves Spirit, mannose, rhamnose, but L-arabinose, maltose, sucrose etc. can not be utilized;Not decomposing protein, it is impossible to which hydrolysis is formed sediment Powder, nitrate is not reduced, does not produce indoles;Catalase reaction negative, negative catalase.Can be containing 6%~8% NaCl rings Grown in border, resistance to NaCl concentration 13%~20%.15-45 DEG C, pH4.5-9.6, pH of growth temperature is given birth to when being subacidity to neutrality Length physically well develops.Pediococcus acidilactici (Pediococcus acidilactici) new strains HT is in preservation on the 04th in 09 month in 2012 In China Committee for Culture Collection of Microorganisms's common micro-organisms center, abbreviation CGMCC, address:The Chaoyang District, Beijing City North Star The institute 3 of West Road 1, Institute of Microorganism, Academia Sinica, deposit number are CGMCC No.6500.
After described Pediococcus acidilactici fermentation, it is freeze-dried, lyophilized bacterium powder can be prepared into, therefore the present invention also carries For the lyophilized bacterium powder of the Pediococcus acidilactici.
The present invention also provides the preparation method of the lyophilized bacterium powder, comprises the following steps:
1) Pediococcus acidilactici HT seed liquor is prepared;
2) seed liquor for preparing step 1) carries out liquid deep layer fermenting culture with 1-3% inoculum concentration;
3) zymotic fluid is centrifuged, concentrated, added freeze drying protectant, vacuum freeze drying, prepare lyophilized bacterium powder.
Wherein, the preparation method of the seed liquor described in step 1) is as follows:The strain HT of test tube slant culture is seeded to MRS In culture medium, 25-37 DEG C of constant-temperatureanaerobic anaerobic culture 16-24h, it is seeded to by 1~5% in seed culture medium, 25-37 DEG C of Anaerobic culturel 16-24h, as seed liquor.
Wherein, the culture parameters of liquid deep layer fermenting culture are in step 2):Temperature, 25-37 DEG C;Mixing speed, 30~ 100rmp;PH value, 6.0 ± 0.5;Incubation time, 16-24h;Fermentation cumulative volume 0.5-1% CaCO is added in fermentation process3, PH is adjusted in 6.2-6.5;The glucose of feed supplement 30~60% is carried out after fermentation 8h.
Bacillus subtilis mushroom bacteriocin freeze-dried powder provided by the invention is prepared as follows:Fermented and cultured is withered Careless bacillus CGMCC No.6511, the class bacteriocin in bacillus subtilis fermentation liquor, dialysis are extracted with ammonium sulfate salting-out process Desalination, freeze-drying obtain concentrated freeze-dried powder.
Lyophilized bacterium powder provided by the invention containing the Pediococcus acidilactici and the bacillus category bacteriocin freeze-dried powder Feed addictive, wherein, the class bacteriocin freeze-dried powder preferably accounts for 40-60%;Pediococcus acidilactici freeze-dried powder preferably accounts for 60- 40%.
Wherein, in the feed addictive, class bacteriocin freeze-dried powder >=100mg/g, viable count of lactobacillus >=1010CFU/g。
Feed addictive provided by the invention, it can be added in conventional livestock and poultry premix, as vitamin and minerals are pre- In batch mixing, therefore the present invention also provides the premix containing the feed addictive.
Further, the feed addictive of the present invention can be added to conventional livestock and poultry basal diet, such as Corn-soybean In type daily ration, or described premix is added in conventional basal diet, is prepared into batch, therefore the present invention also provides Batch containing the feed addictive.
The class bacteriocin of the present invention and Pediococcus acidilactici mixed fodder additive, it is possible to reduce or avoid breeding production moderate resistance The use of raw element, medicament residue is reduced and eliminated afterwards, improves meat, eggs and milk product quality problem, improves enteric microorganism environment, Preventing and treating diarrhoea produces, and improves production performance, improves feed efficiency, reduces production cost, improves immunity and premunition.
The class bacteriocin of the present invention and Pediococcus acidilactici mixed fodder additive, take full advantage of class bacteriocin and lactic acid sheet The advantages of coccus:
(1) Pediococcus acidilactici is safe and effective as one kind, and one of widely used lactic acid bacteria, acid producing ability is strong, can compete Property restrain pathogenic microorganism, adjust gastrointestinal bacterial flora, so as to keep intestinal microecology balanced, be immunized to strengthening animal body Effect has certain effect.
(2) Pediococcus acidilactici acid-fast ability is strong, and bile tolerance ability is strong, and can illustrate that it has to poor environment with high temperature resistant Very strong tolerance, stability is good in feed manufacturing, storage and alimentary canal, therefore can extensive use.
(3) bacillus subtilis mushroom bacteriocin has broad spectrum antibacterial, can suppress E.coli K88, e. coli k99, Bacterium enteritidis, salmonella typhimurium, and aspergillus niger and aspergillus flavus, there is stronger heat endurance, intestines can be promoted The beneficial microorganism growing multiplications such as road endospore bacillus, lactic acid bacteria and saccharomycete and metabolism is enlivened, and activity is high.Cause This, class bacteriocin is combined with lactic acid bacteria club's performance effect or effect considerably beyond single use effect.
(4) product of the present invention can be widely applied to pig, fowl, ox, sheep, aquatic products and economic animal, be a kind of green feed Additive, there are economic benefit and ecological benefits.
Bacillus subtilis (Bacillus subtilis) in the present invention separates for applicant from traditional zymotic fermented soya bean Obtain, its biological property is as follows:Individual cells, 0.7-0.8 × 2-3 μm of bacterium colony rough surface is opaque, dirty white or micro- Huang Color, bacterium colony is circular, and edge is serrated, gram-positive bacteria;0.6-0.9 × 1.0-1.5 μm of gemma, form arrive post for ellipse Shape, central or slightly inclined positioned at thalline, thalline does not expand after sporulation.Aerobic bacteria.Bacillus subtilis in the present invention has Beneficial effect is:
(1) the bacillus subtilis acid resistance in the present invention is strong, bile tolerance, high temperature resistant (tolerable 80 DEG C granulation temperature Degree), the bioactivity that changes can be kept after animal intestinal tract is entered afterwards, effectively suppress enteropathogenic E. Coli K88, K99 and Staphylococcus aureus, promote beneficial to bacteria growing.
(2) can thalline itself synthesis alpha-amylase, protease, lipase, the enzyme such as cellulase, in alimentary canal with Digestive enzymes in animal body together plays a role, it is possible to increase feed nutrient digestibility, improves breeding performonce fo animals, is advantageous to Reduce culture-cycle and aquaculture cost.
(3) a variety of B family vitamins such as vitamin B1, B2, B6, nicotinic acid can be synthesized, are normal growth and the life of animal body The performance of reason function provides safeguard, and improves the activity of interferon and macrophage in animal body, strengthens immunity function and anti-disease Sick ability.
The bacillus subtilis (Bacillus subtilis) of the present invention is preserved in the micro- life of China on the 4th in September in 2012 Thing culture presevation administration committee common micro-organisms center, abbreviation CGMCC, address:Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Number, Institute of Microorganism, Academia Sinica, deposit number is CGMCC No.6511.
Embodiment
Following examples are used to illustrate the present invention, but are not limited to the scope of the present invention.
The preparation of the bacillus subtilis CGMCC No.6511 class bacteriocins of embodiment 1
1) preparation of bacillus subtilis fermentation liquor
It is 7 to carry out fermentation 20h, initial pH using MNB culture mediums under 30 DEG C of cultivation temperatures, inoculum concentration 3%, shaking speed 225rpm。
The formula of MNB culture mediums is:Glucose, 5.0g/L;Peptone, 5.0g/L;Meat extract, 3.0g/L;Yeast extract, 1.0g/L;MgSO47H2O, 0.5g/L;MnSO4·H2O, 0.005g/L;PH, 7.0.
2) ammonium sulfate salting-out process extraction class bacteriocin
Cultured bacillus subtilis bacterium solution is prepared into bacteria-free filtrate, ground ammonium sulfate powder is added slowly to In bacteria-free filtrate, and it is gently agitated for ensuring that ammonium sulfate is completely dissolved, after slowly stirring 1h, is positioned in 4 DEG C of refrigerators.Ammonium sulfate Gradient processing is 70%.After overnight, the solution of different saturation is taken out, centrifuges (10000 × g, 30min, 4 DEG C) respectively, is received Collection precipitation.
3) dialysis desalting
By the precipitation of collection, 20mMTris-HCl cushioning liquid (pH 8.01) is dissolved in, loads retention relative molecular mass In 3500Da bag filter (Amersham, Sweden), after being dialysed in the Tris-HCl cushioning liquid of same concentrations, with carrying out Freeze-drying obtains concentrated dried powder, 4 DEG C of preservations.
4) measure of protein concentration
Class bacteriocin obtained by preparing is subjected to protein concentration using BCA kits (Pierce, Rockford, IL) Measure, sample are added on ELISA Plate, and absorbance is determined at 570nm, and egg is calculated according to standard appended by kit and computational methods White matter content, the μ g/mL of class bacteriocin protein concentration 57.1 extracted when 70%.
5) measure of antibacterial activity
1mg/mL is arrived into the regulation of obtained class bacteriocin concentration, it is determined to Pediococcus acidilactici, golden yellow using cylinder plate method Staphylococcus, Bacterium enteritidis, E.coli K88, the antibacterial activity of e. coli k99 and salmonella typhimurium, are treated pair After covering with flat board according to (20mM Tris-HCl buffer solutions) lawn, the antibacterial circle diameter (d) of antibacterial thick protein, each place are measured Manage 3 repetitions.
The Determination of Antibacterial Activity of the bacillus subtilis mushroom bacteriocin material of table 1
Indicator bacteria Culture medium Temperature (DEG C) Inhibition zone (mm)
Pediococcus acidilactici CGMCC No.6500 MRS 37 0
Staphylococcus aureus IVDC C56005 BHI 30 31
Bacterium enteritidis IVDC C79-52 BHI 37 20
Escherichia coli k88IVDC C83901 BHI 30 23
Escherichia coli k99IVDC C83529 BHI 30 27
Salmonella typhimurium IVDC C77-31 BHI 37 22
As can be seen from Table 1, the class bacteriocin of producing bacillus subtilis life selects us the Pediococcus acidilactici of addition There is no inhibitory action, to pathogenic bacteria staphylococcus aureus, Bacterium enteritidis, E.coli K88, e. coli k99 and mouse The growth and breeding inhibitory action of salmonella typhi is stronger.These pathogens are all the Main Pathogenic Bacterias of animal in livestock-raising.
The preparation of the Pediococcus acidilactici bacterium powder of embodiment 2
1) preparation of Pediococcus acidilactici CGMCC No.6500 seed liquors
Test tube slant preservation of bacteria strain is seeded in the anaerobism pipe equipped with 20mL MRS fluid nutrient mediums, 37 DEG C of constant-temperatureanaerobic anaerobics 20h is cultivated, treats viable count up to 107CFU/mL, the 250mL conical flasks equipped with 100mL MRS fluid nutrient mediums are inoculated in by 1% In, 37 DEG C of Anaerobic culturel 20h, treat viable count up to 107More than CFU/mL (MRS agar mediums plate count), as seed liquor It is standby.
2) liquid deep layer fermenting culture
Above-mentioned Pediococcus acidilactici liquid seeds liquid is added in 50L fermentation tanks with 1% inoculum concentration, culture parameters are: Temperature, 37 DEG C;Mixing speed, 50rmp;PH value, 6.0 ± 0.5;Incubation time, 20h.Fermentation cumulative volume is added in fermentation process 0.5% CaCO3, pH is in 6.2-6.5 for regulation;Feed supplement is carried out after fermentation 8h, that is, adds 50% glucose 1L.
The MRS Liquid Culture based formulas cultivated used in Pediococcus acidilactici is as follows:Peptone, 10.0g/L;Beef extract, 8.0g/ L;Yeast extract, 4.0g/L;Glucose, 20.0g/L;Tween 80,1mL/L;Dipotassium hydrogen phosphate, 2.0g/L;Sodium acetate trihydrate, 5.0g/L;Triammonium citrate, 2.0g/L;Epsom salt, 0.2g/L;Manganese sulfate, 0.05g/L;pH 6.2±0.2.
MRS agar mediums are that agar 15g/L is added on the basis of MRS fluid nutrient mediums.
Optimization culture based formulas used in 50L fermentation tank Multiplying culture Pediococcus acidilacticis is as follows:Peptone, 10.0g/L;Ox Meat extract, 8.0g/L;Yeast extract, 6.0g/L;Glucose, 25.0g/L;Tween 80,1mL/L;Dipotassium hydrogen phosphate, 3.0g/L;Three water Sodium acetate, 5.0g/L;Triammonium citrate, 2.0g/L;Epsom salt, 0.2g/L;Manganese sulfate, 0.05g/L;pH 6.2±0.2.
3) bacterium powder is freezed to prepare
By the centrifugation of zymotic fluid, concentration, freeze drying protectant is added (containing 5% trehalose, 15% skimmed milk power, 4% sugarcane Sugar, 1%D- arabo-ascorbic acids and 2.0% sodium glutamate), vacuum freeze drying and bacterium powder viable count the production routine such as examine, most Powdery active bacteria formulation is prepared eventually.
4) finished product detection
The powdery active bacteria formulation 1g prepared is taken, is determined by international GB/T 4789.35-2003 methods in every gram of bacterium powder Viable count, and viable count is more than 10 in bacterium powder8CFU/g。
The preparation of the class bacteriocin of embodiment 3 and Pediococcus acidilactici mixed fodder additive
Lactic acid prepared by the bacillus category bacteriocin dry powder 50% (wt) being prepared in embodiment 1 and embodiment 2 Piece coccus bacterium powder 50% (wt) is well mixed to be formed.Its quality standard is:Class bacteriocin dry powder 102mg/g, viable count of lactobacillus 12000000000/g, moisture≤10%, arsenic≤0.5mg/kg, lead≤1.0mg/kg.
Test example 1
The application effect that class bacteriocin of the present invention grows with Pediococcus acidilactici mixed fodder additive to piglet
1st, experimental animal
Du × length × Dasanyuan healthy growth pig 72 that average original body mass is 23.12 ± 1.58kg.
2nd, experimental design
By the close principle similar with hereditary basis of body weight, three processing are randomly divided into, each handle 6 repetitions, each Repeat (circle) 4 pigs, male and female half and half, experimental period 20-50kg.Group one is blank control group, feeds basal diet;Group 2 is anti- Raw plain control group, basal diet addition 100g/t aureomycin, 50g/t flavomycoins;Group three is test group, is added in basal diet The mixed fodder additive that 0.2% embodiment 3 is prepared.
3rd, daily ration is tested, is shown in Table 2
Table 2 tests Diet Formula and nutrient component meter
4th, feeding management
Stable breeding, free choice feeding and drinking-water, inventory and residual feed and abnormal conditions are recorded daily, in off-test pair Pig is weighed.Routinely program and method are immunized.
5th, test results and analysis
Compared with blank control group, the indices improvement of test group is as follows:Average daily gain improves 10.7%, averagely Daily gain improves 12.1%, and feed conversion rate improves 13.0%, and diarrhea rate reduces by 71.2%;Compared with antibiotic control group, examination The indices improvement for testing group is as follows:Average daily gain improves 9.3%, and average daily gain improves 10.8%, feed conversion rate Improve 12.3%, diarrhea rate reduces by 56.5%.Have the above results it should also be appreciated that, addition the present invention in mixing premix growth-promoting Effect that is long and reducing diarrhea rate is better than the result obtained by addition antibiotic.
Test example 2
The influence of class bacteriocin of the present invention and Pediococcus acidilactici mixed fodder Additive on Production Performance of Broilers
1st, experimental animal and design
Laid eggs the blue laying hen 180 in later stage sea from age in days identical, be divided into three groups, every group of 10 repetitions are each to repeat 6 Chicken.Group one is blank control group, feeds basal diet;Group 2 is antibiotic control group, and basal diet adds 100g/ aureomycin, 50g/ flavomycoins;Group three is test group, and the mixed fodder additive that 0.2% embodiment 3 is prepared is added in basal diet.2、 Test daily ration
Feed formulation prepares corn-soybean meal basal diet with reference to NRC (1994) broiler chicken nutrition requirements.Each processing base Plinth Diet Formula and trophic level are shown in Table 3.All daily rations are powder.
Table 3 tests Diet Formula and nutrient component meter
3rd, test results and analysis
As can be seen from Table 4,0-21 days stages, 22-42 days stages and 0-42 days, average day feeding between three groups Notable difference is not present in amount, average daily gain, and only feed weightening ratio has significant difference.With group two i.e. antibiotic control group phase Than, for three mixed fodder additive groups of group at 0-21 days, 22-42 days, 0-42 days, feed weightening ratio was respectively increased 14.1%, 14.06%, 9.39%.Though significant difference is not present between two blank control groups of group three and group, compared with, three Broiler chicks of group are raised Material weightening is than still there is the trend of reduction.Illustrate that this product can improve the production performance of Broiler chicks, Er Qiewu to a certain extent Poison, have no drug resistance, it is green safe, useful external source probio is not only with the addition of to animal body, the class bacteriocin of addition can also Promote the growth of flora in animal intestinal tract, improve feed nutrient digestion and absorption so as to realize, improve immune and premunition work( Effect, and the function of antibiotic usage is reduced and eliminates, permanent use can produce economic benefit, social benefit and Ecological Effect Benefit.
The influence of the mixed fodder Additive on Production Performance of Broilers of table 4
Note:A, b without same letter are significant difference (P with the shoulder mark of a line<0.05);SEM misses for average.
Test example 3
The influence that class bacteriocin of the present invention cultivates with Pediococcus acidilactici mixed fodder additive to grass carp
From 9000, grass carp, it is divided into three treatment groups, group one is blank control group (basal diet group);Based on group two Daily ration+100g/t aureomycin+100g/t flavomycoins;The mixed fodder of daily ration+0.2% additive (is made by embodiment 3 based on group three It is standby), 3 repetitions are each handled, it is each to repeat 1000.Compared with group one, three feed coefficients of group reduce by 8.2%, averagely increase day by day Bring up again high by 12.6%, survival rate raising 23.7%;Compared with group two, three feed coefficients of group reduce by 7.3%, and average daily gain improves 11.8%, survival rate improves 20.9%, and the regularity of group three is good, and resistance against diseases is strong.In terms of Water-quality control, 3 meters, 3 mu of the depth of water Launch the mixed fodder additive 1kg that embodiment 3 obtains.As a result in 24h, in water ammonia nitrogen concentration by group two 0.72mg/L The 0.21mg/L of test group is reduced to, reduction three times are more, illustrate that product of the present invention can only not improve aquaculture production effect The effect of rate, additionally it is possible to purify the growing environment of aquatic livestock, improve water quality.
Described above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art For member, without departing from the technical principles of the invention, some improvements and modifications can also be made, these improvements and modifications Also it should be regarded as protection scope of the present invention.

Claims (8)

1. a kind of feed addictive, it is characterised in that contain the Pediococcus acidilactici that deposit number is CGMCC No. 6500 (Pediococcus acidilactici)Lyophilized bacterium powder and bacillus subtilis CGMCC No. 6511 class bacteriocin freeze Dry powder.
2. feed addictive as claimed in claim 1, it is characterised in that the Pediococcus acidilactici freezes the preparation side of bacterium powder Method, comprise the following steps:
1)Prepare the seed liquor of the Pediococcus acidilactici;
2)By step 1)The seed liquor of preparation carries out liquid deep layer fermenting culture with 1-3% inoculum concentration;
3)Zymotic fluid is centrifuged, concentrated, freeze drying protectant is added, vacuum freeze drying, prepares lyophilized bacterium powder.
3. feed addictive as claimed in claim 2, it is characterised in that step 1)The preparation method of described seed liquor is such as Under:The strain HT of test tube slant culture is seeded in MRS culture mediums, 25-37 DEG C of constant-temperatureanaerobic anaerobic culture 16-24h, connect by 1 ~ 5% Kind is into seed culture medium, 25-37 DEG C of Anaerobic culturel 16-24h, as seed liquor.
4. feed addictive as claimed in claim 2, it is characterised in that step 2)The culture ginseng of middle liquid deep layer fermenting culture Number is:Temperature, 25-37 DEG C;Mixing speed, 30 ~ 100rmp;PH value, 6.0 ± 0.5;Incubation time, 16-24h;In fermentation process Add the 0.5-1% of fermentation cumulative volume CaCO3, pH is in 6.2-6.5 for regulation;The glucose of feed supplement 30 ~ 60% is carried out after fermentation 8h.
5. feed addictive as claimed in claim 1, it is characterised in that the bacillus subtilis mushroom bacteriocin freeze-dried powder is pressed Prepared according to following manner:Bacillus subtilis described in fermented and cultured, sent out with ammonium sulfate salting-out process extraction bacillus subtilis Class bacteriocin in zymotic fluid, dialysis desalting, freeze-drying obtain concentrated freeze-dried powder.
6. feed addictive as claimed in claim 1, it is characterised in that the class bacteriocin freeze-dried powder accounts for 40-60%;It is described Pediococcus acidilactici freeze-dried powder accounts for 60-40%.
7. the premix containing the feed addictive described in claim 1 or 6.
8. the batch containing the feed addictive described in claim 1 or 6.
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