CN104498385B - High yield antibacterial peptide Bacillus licheniformis and application thereof - Google Patents
High yield antibacterial peptide Bacillus licheniformis and application thereof Download PDFInfo
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- 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
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- 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
- C12R2001/10—Bacillus licheniformis
Abstract
Does is the invention discloses a kind of Bacillus licheniformis, Bacillus licheniformis (Bacillus? licheniformis) ZJU-X1, is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, does is preserving number CGMCC? No.9688.The present invention also also discloses and utilizes above-mentioned Bacillus licheniformis to prepare and the Bacillus licheniformis preparation that obtains, and this Bacillus licheniformis preparation is as fodder additives.The present invention also provides simultaneously and utilizes above-mentioned Bacillus licheniformis to prepare and the animal microorganism preparation that obtains, the microbiotic of this animal microorganism preparation alternatively in weaning pigs; In described animal microorganism preparation, the viable count of Bacillus licheniformis preparation is 10
11cfu/g.
Description
Technical field
The present invention relates to fodder additives---microbial preparation field, is specifically related to a kind of Bacillus licheniformis and application thereof of highly effective and safe.
Background technology
Feeding antibiotic has significant effect in promotion growth of animal, raising feed efficiency, the loss of minimizing grain, raising pork output etc.But along with microbiotic is as Animal feed-additive widely using in feed; its drawback shows especially just day by day; as caused animal gastrointestinal tract flora imbalance, producing resistance and drug residue etc., having caused the problem of a series of public safety, environment protection and animal products quality aspect.Find green safety fodder additives that is nontoxic, that efficiently, do not produce resistance and become one of Animal nutrition field focus.
Bacillus licheniformis (bacilluslicheniformins) is one of feed grade bacterial strain of the Ministry of Agriculture of China No. 318 bulletin approval uses in 2003.It is the amphimicrobe that can produce gemma.Taxonomy belongs to prokaryotic organism, eubacterium.Because it has fast, the easy cultivation of thalline reproduction speed, strong stress resistance, high temperature resistant, acid and alkali-resistance, anti-extrusion, and the features such as energy extracellular proteinase, amylase, lipase and multiple amino acids, so the wide model of its product application scope, particularly in medicine and livestock-raising, there is higher using value.
Bacillus licheniformis (bacilluslicheniformins) plays a role mainly through the following aspects: 1. maintain the microecological balance in animal intestinal, promote propagation and the growth of probiotics (lactobacillus, bifidus bacillus), suppress the growth and breeding of the harmful bacterias such as harmful bacteria (intestinal bacteria, Salmonellas).2. genus bacillus in the formulation within raw spore form exist.After spore enters animal intestinal, can bring back to life rapidly in upper intestines proteolytic enzyme, lipase, the amylase that also secretion activity is very strong, contribute to the carbohydrate of some complexity in degrading plant feed, produce the many kinds of substance with antagonism pathogen enterobacteria.3. cause anaerobic environment by the biological oxygen taken by force in oxygen consumption enteron aisle, enteron aisle ancestral home dominant bacteria belongs to anerobe mostly, and harmful bacteria and foreign bacteria mostly are aerophil, thus is conducive to maintaining intestinal eubiosis.4. strengthen the non-specific immunity of animal, resist infection.
The domestic correlative study to Bacillus licheniformis report is few, and research mainly concentrates on zymotechnique and makes on product, less to its applied research in growth of animal.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of Bacillus licheniformis and application thereof.
In order to solve the problems of the technologies described above, the invention provides a kind of Bacillus licheniformis, for Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCCNo.9688.
The present invention also provides simultaneously and utilizes above-mentioned Bacillus licheniformis to prepare and the Bacillus licheniformis preparation that obtains: form by as Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1 of effective constituent and carrier;
Described carrier is at least one in maltodextrin, wheat bran, dregs of beans, stone flour, Semen Maydis powder;
Described Bacillus licheniformis preparation is as fodder additives.
The present invention also provides simultaneously and utilizes above-mentioned Bacillus licheniformis to prepare and the animal microorganism preparation that obtains: comprise Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1 as activeconstituents I and carrier;
Described carrier is at least one in Semen Maydis powder, maltodextrin, wheat bran, dregs of beans, stone flour;
The microbiotic of described animal microorganism preparation alternatively in weaning pigs.
Improvement as animal microorganism preparation of the present invention: animal microorganism preparation also comprises active component II;
Described active component II is at least one in clostridium butylicum, subtilis, milk-acid bacteria, bifidus bacillus.
Improvement as animal microorganism preparation of the present invention: in animal microorganism preparation, the viable count of Bacillus licheniformis preparation is 10
11cfu/g.
Bacterial strain of the present invention is separated from health pig enteron aisle, belong to gemma Cordycepps, bacillus, Gram-positive, cellular form and arrangement are in shaft-like, Dan Sheng, produce nearly middle raw ellipticity gemma, sporangiocyst slightly expands, high temperature resistant, acidproof, alkaline-resisting, produce enzyme, secretion antibacterial peptide also has the strain characteristic of microbial balance regulating animal intestinal, this bacterial strain ZJU-X1 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation title: Bacillus licheniformis (Bacilluslicheniformis), preservation date is on September 17th, 2014, preserving number is CGMCCNo.9688, address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode 100101.
Of the present invention the provided application of Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1 in following at least one also belongs to protection content of the present invention:
1. Bacillus licheniformis preparation is prepared;
2. fodder additives probiotics preparation is prepared;
2. antibacterial peptide is prepared;
4. animal-feed is prepared;
5. the microbiotic in feed is substituted;
6. the growth of pig is promoted.
The present invention is by being separated, identifying, screen, obtain Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1, its strong stress resistance, there is prebiotic function, the present invention is made into Bacillus licheniformis preparation, prepare animal-feed as fodder additives, wherein animal includes but not limited to the various animals such as pig.This additive has the use of alternative fodder antibiotics, regulates animal intestinal micro-ecology balance, strengthens the effect of animal non-specific immunity, produce the nutritional factor such as VITAMIN, useful enzyme simultaneously, promotes that animal digestion absorbs, reduces diarrhoea, growth promoting effects.Bacillus licheniformis of the present invention does not produce resistance, does not remain in animal body, and can not produce human health and threaten, be healthy and safe fodder additives.
Accompanying drawing explanation
Below in conjunction with accompanying drawing, the specific embodiment of the present invention is described in further detail.
Fig. 1 be Bacillus licheniformis ZJU-X1 colonial morphology (bacterium colony be flat, edge is irregular, white, surface irregularity gauffer);
Fig. 2 is Bacillus licheniformis ZJU-X1 gramstaining schematic diagram (this bacterial strain Gram-positive bacillus, therefore gramstaining result is bluish voilet, cellular form and arrangement are in shaft-like, Dan Sheng, and produce ellipticity gemma raw closely, sporangiocyst slightly expands);
Fig. 3 is the growth characteristics of Bacillus licheniformis and produces proteolytic enzyme performance chart;
Fig. 4 is the growth characteristics of Bacillus licheniformis and produces amylase performance chart;
Fig. 5 is the electrophorogram of the antibacterial peptide of Bacillus licheniformis secretion;
Fig. 6 is the thermotolerance comparison diagram of bacillus licheniformis.
Embodiment
The experimental technique used in following case study on implementation if no special instructions, is ordinary method.
The experiment material used in following case study on implementation, reagent etc., if no special instructions, all obtain by commercial channel.
Below in conjunction with concrete case study on implementation, the present invention is described in further detail.
The separation of embodiment 1 bacterial classification, qualification and characteristic research
1, experimental technique
1.1 pig farm, experiment material Zhejiang Province sodium selenite intestinal contents 10 parts.
1.2 substratum
1. count substratum (being mass concentration g/L): peptone 10g, beef extract 5g, sodium-chlor 5.0g, agar 15.0g, distilled water is settled to 1.0L, pH7.2,121 DEG C, 15min sterilizing.
2. LB substratum (being mass concentration g/L): peptone 20g, K
2hPO
4.3H
2o2.5g, MgSO
40.73g, glycerine 15.0ml, agar 15.0g, distilled water is settled to 1.0L; PH7.0,121 DEG C, 15min sterilizing.
1.3 specific experiment methods
1) the intestinal contents 10g getting faecal samples sodium selenite is added in 90mLPBS damping fluid, puts into 80 DEG C of water-bath 10min; Thus kill non-sporeformer, then put into 20mlLB liquid nutrient medium, cultivate 24h in 37 DEG C;
2) sample is diluted to 10 respectively
-3, 10
-5, 10
-7, coat LB solid medium, put into constant incubator, cultivate 24h for 37 DEG C;
3) adopt replica-plating method to be separated bacterial classification, choose well-grown, to have radial fringe bacterial strain, carry out microscopy;
4) choose the bacterial strain that cultural characteristic, colonial morphology and microscopic morphology conform to Bacillus licheniformis and carry out Physiology and biochemistry qualification.
5) 16SrDNA order-checking qualification
Gene sequencing adopts colony polymerase chain reaction (PCR) method amplification to obtain 16SrDNA fragment.Bacteria total DNA is extracted and is adopted UNIQ-10 pillar bacterial genomes DNA extraction agent box (the raw work in Shanghai, SK8229) to extract.16SrDNA amplimer adopts bacterium amplification by primer, and forward primer is 7F:5 ’ – CAGAGTTTGATCCTGGCT – 3 '; Reverse primer is 1540R:5 ’ – AGGAGGTGATCCAGCCGCA-3 '.Reaction system (25 μ L): 5 × Buffer (withMg
2+) 2.5 μ L; DNTP (each 2.5mM) 1 μ L; 7F (10uM/ μ L) 0.5 μ L; 1540R (10uM/ μ L) 0.5 μ L; Genomic dna (50ng/ μ l) 0.5 μ L; TaKaRaExtaq enzyme (5U/ μ L) 0.3 μ L; Add distilled water to 25 μ L.Pcr amplification program: 98 DEG C of denaturation 3min, 98 DEG C of sex change 25sec, 55 DEG C of annealing 25sec, 72 DEG C extend 1min, carry out 30cycle, and last 72 DEG C extend 4min.
PCR primer utilizes 1% agarose gel electrophoresis to detect, and serves the raw work sequencing in sea after the positive products of fragment 1500bp is purified.
6), amylase, protease activity determination and high temperature resistant determination of activity is produced.
2, experimental result
2.1 Morphological Identification
After 10 parts of sample preparation are cultivated, constant temperature culture is carried out with photocopy flat board, obtain bacterial strain 8 strain, selective medium is cultivated rear mirror mirror and is obtained lichem bacillus strain 1 strain, and be initially identified as Bacillus licheniformis after colonial morphology qualification and gram stain microscopy after carrying out purifying, its diameter is 0.6-0.8 × 1.5-3.5 μm, end circle, there is gemma, have peritrichous, can move.(see Fig. 1 and Fig. 2).
2.2 physiological and biochemical property
The method introduced in basis " qualification that bacterium belongs to is instructed " subsequently, mensuration isolate is to the utilization power of various carbon source and carry out other physiological and biochemical tests.The results are shown in Table 1.
Table 1 is separated the physiological and biochemical property of Bacillus licheniformis
| Project name | Result | Project name | Result |
| Glucose | + | Gram-positive | + |
| Pectinose | + | Gemma | + |
| Sucrose | + | Catalase | + |
| Seminose | + | Sports type | + |
| Zulkovsky starch | + | Anaerobic growth | - |
| Wood sugar | + | 3%NaCl grows | + |
| Fructose | + | Nitrate reduction | + |
| Rhamnosyl | - | ||
| Sorbyl alcohol | + | ||
| Trehalose | + |
Note :+represent that result is positive ,-represent that result is negative
According to test-results, contrast " uncle Jie Shi Bacteria Identification handbook " retrieval, bacterial strain preliminary evaluation result is Bacillus licheniformis.2.316SrDNA sequence homology analysis.
Sequence 1, as shown in SEQIDNO:1 (sequence 1), is carried out BLAST (http://blast.ncbi.nlm.nih.gov/Blast.cgi) sequence analysis at Genbank database, is defined as Bacillus licheniformis by its sequence.
2.4 cultural characterss and the analysis of product digestive ferment
(namely the mono-clonal of the Bacillus licheniformis ZJU-X1 through sequence verification is inoculated in LB substratum, the Bacillus licheniformis ZJU-X1 getting 1 transfering loop is inoculated in the LB substratum of 5ml), after 37 DEG C of constant temperature culture cultivate 24h, utilize amylase, proteolytic enzyme detection kit (Bioisystech Co., Ltd is built up in Nanjing) carries out digestive enzyme activity detection.The results are shown in Figure 3 and Fig. 4.
According to Fig. 3 and Fig. 4, learn: Bacillus licheniformis ZJU-X1 has higher product amylase and the activity of proteolytic enzyme, after cultivation 48h, the activity of producing amylase and proteolytic enzyme reaches the highest.
2.5 antibacterial peptide ability qualifications
(namely the mono-clonal of the Bacillus licheniformis ZJU-X1 through sequence verification is inoculated in LB substratum, the Bacillus licheniformis ZJU-X1 getting 1 transfering loop is inoculated in the LB substratum of 5ml), cultivate different time gradient (37 DEG C of constant-temperature tables, 220rpm, cultivate 8h, 16h, 24h, 32h, 40h, 48h, 56h) after, the centrifugal 5min of 1000rpm, gets supernatant, carry out electrophoresis checking, as described in Figure 5.In Fig. 5, be followed successively by 8h, 16h, 24h, 32h, 40h, 48h, 56h time of cultivation from left to right.According to Fig. 5, we learn: after cultivation 24h, the output of antibacterial peptide is the highest.And after ion-exchange and sieve chromatography purifying, obtain antibacterial peptide, and verify.
Simultaneously, by identical method, to the wild-type Bacillus licheniformis bought from Chinese Organism Depositary, (CGMCC numbers: product antibacterial peptide ability 1.265) measures for we, result shows, its antibacterial peptide production capacity is significantly lower than Bacillus licheniformis ZJU-X1.
Remarks illustrate: above-mentioned ion-exchange and sieve chromatography purification process, and the qualification of antibacterial peptide ability, can refer to invention " antibacterial peptide of a kind of Bacillus licheniformis secretion and its preparation method and application the " (application number: 2014100219061 published; Application publication number: CN103772490A) carry out.
2.6 thermotolerance outcome researches
The mono-clonal of the Bacillus licheniformis ZJU-X1 through sequence verification is inoculated in LB substratum, after cultivating 24h, get culture centrifugal, Eddy diffusion after 0.9% brine twice, be placed in 60 DEG C respectively, 70 DEG C, 80 DEG C, 90 DEG C, 100 DEG C of temperature water bath process 15 minutes, rapid flowing water cooling, then adopts dilution-plate method to measure genus bacillus survival rate, the results are shown in Figure 6.Result shows, this bacterial strain, 90 DEG C time, still has higher survival rate, has good thermotolerance.
This bacterial strain is Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, preserving number is CGMCCNo.9688, preservation date on September 17th, 2014.
The application of embodiment 2 Bacillus licheniformis substitute antibiotics in weanling pig
1. object and meaning: select weanling pig to be subjects, inquires into Bacillus licheniformis under antibiotic-free, condition to the impact of Growth Performance of Weaning Piglets, diarrhea rate.
2. materials and methods:
Experimental animal: choose 23 ± 2 days age in days Du × length × Dasanyuans and hybridize healthy weanling pig 60, body weight is 7.8 ± 0.2Kg.
Bacillus licheniformis preparation (animal microorganism preparation): through drying gained after Bacillus licheniformis ZJU-X1 liquid fermenting, Semen Maydis powder is carrier, and the viable count of obtained Bacillus licheniformis preparation is 10
11cfu/g, in the implementation case, this Bacillus licheniformis preparation uses as animal additive.
Remarks illustrate: processing step and the parameter of above-mentioned Bacillus licheniformis ZJU-X1 liquid fermenting are: by 1% inoculum size inoculation seed liquor in nutrient solution, in 220rpm, cultivate 48 hours at 37 DEG C; After end by bacterium liquid even spraying on Semen Maydis powder, and stirring and evenly mixing, is placed on 60 DEG C of oven for drying to moisture <10%.
Test grouping: by healthy for 60 ages weanling pig, according to body weight, nest not, sex is divided into 2 groups, establishes three repetitions for each group, each repetition 10,14 days trial periods.Process 1 is control group (microbiotic positive controls), basal diet+and microbiotic (olaquindox 100ppm+ sulphur glues 200ppm+ kitasamycin 100ppm); Process 2 is experimental group, basal diet+Bacillus licheniformis preparation 10
9cfu/T.NRC2012 standard formulation pressed by basal diet.
3. feeding animals management
Test weanling pig adopts high bed to raise, natural ventilation, and environmental health condition is good, free choice feeding, drinking-water; Expelling parasite and immunity are according to pig farm code process.Observe feed intake and health condition at any time.All diarrhoea pigs are not treated.
4. testing index
The mensuration of 4.1 growth performances
Test and when starting and when terminating, piglet and feed are weighed, calculate the average daily gain of every pig of each treatment group and average food consumption, calculate feedstuff-meat ratio.Feedstuff-meat ratio is the ratio of per day food consumption and per day weightening finish.
4.2 diarrhea rates measure
In whole process of the test, every morning 9 observes diarrhea of pigs situation, and every pig is only added up once every day.Diarrhea rate is the ratio of diarrhoea piglet head number and the total head number of piglet.
5. data statistics: test all single factor test methods analysts occupying SPSS17.0 (SPSSInc., USA) statistical software of searching for food and carry out significance analysis, significant difference then carries out DunanShi method multiple comparison analyse.Data mean value ± standard error represents.
6. experimental result
Table 2 Bacillus licheniformis ZJU-X1 substitutes antibiotic research in creep feed
Table 2
| Control group | Experimental group | |
| Starting weight/Kg | 7.73±0.07 | 7.76±0.03 |
| End weight/Kg | 9.37±0.03 | 9.47±0.08 |
| ADG/g (day weight gain) | 137.12±7.95 | 142.05±4.21 |
| ADFI/g (daily ingestion amount) | 275.94±6.97 | 265.89±5.54 |
| G:F/g/kg (feed-weight ratio) | 496.44±20.18 | 534.47±15.90 |
| Diarrhea rate/% | 14.77±1.31 | 11.36±0.66 |
The original body mass of each test group weanling pig, day weight gain, feed-weight ratio and diarrhea rate are in table 2.
As seen from the experiment, add Bacillus licheniformis preparation in daily ration, the weanling pig speed of growth comparatively treatment group improves, and feed conversion rate improves, and illustrates that Bacillus licheniformis can replace the antibiotic usage in weaning pigs completely.Bacillus licheniformis preparation can show the diarrhoea situation of low weanling pig.
Can obtain from experimental result, this Bacillus licheniformis preparation is at weanling pig stage complete alternative microbiotic.
Remarks illustrate: Bacillus licheniformis ZJU-X1 and clostridium butylicum compound action effect, can see the experiment effect of patent " a kind of clostridium butylicum and application thereof ".
Finally, it is also to be noted that what enumerate above is only several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be had.All distortion that those of ordinary skill in the art can directly derive from content disclosed by the invention or associate, all should think protection scope of the present invention.
<110> Zhejiang University
<120> high yield antibacterial peptide Bacillus licheniformis and application thereof
<160>1
<210>1
<211>1468
<212>DNA
<213> Bacillus licheniformis (
bacilluslicheniformis)
<400>1
gtgctatacatgcagtcgagcggacagatgggagcttgctccctgatgttagcggcggac60
gggtgagtaacacgtgggtaacctgcctgtaagactgggataactccgggaaaccggggc120
taataccggatgcttgattgaaccgcatggttcaattataaaaggtggcttttagctacc180
acttacagatggacccgcggcgcattagctagttggtgaggtaacggctcaccaaggcaa240
cgatgcgtagccgacctgagagggtgatcggccacactgggactgagacacggcccagac300
tcctacgggaggcagcagtagggaatcttccgcaatggacgaaagtctgacggagcaacg360
ccgcgtgagtgatgaaggttttcggatcgtaaaactctgttgttagggaagaacaagtac420
cgttcgaatagggcggtaccttgacggtacctaaccagaaagccacggctaactacgtgc480
cagcagccgcggtaatacgtaggtggcaagcgttgtccggaattattgggcgtaaagcgc540
gcgcaggcggtttcttaagtctgatgtgaaagcccccggctcaaccggggagggtcattg600
gaaactggggaacttgagtgcagaagaggagagtggaattccacgtgtagcggtgaaatg660
cgtagagatgtggaggaacaccagtggcgaaggcgactctctggtctgtaactgacgctg720
aggcgcgaaagcgtggggagcgaacaggattagataccctggtagtccacgccgtaaacg780
atgagtgctaagtgttagagggtttccgccctttagtgctgcagcaaacgcattaagcac840
tccgcctggggagtacggtcgcaagactgaaactcaaaggaattgacgggggcccgcaca900
agcggtggagcatgtggtttaattcgaagcaacgcgaagaaccttaccaggtcttgacat960
cctctgacaaccctagagatagggcttccccttcgggggcagagtgacaggtggtgcatg1020
gttgtcgtcagctcgtgtcgtgagatgttgggttaagtcccgcaacgagcgcaacccttg1080
atcttagttgccagcattcagttgggcactctaaggtgactgccggtgacaaaccggagg1140
aaggtggggatgacgtcaaatcatcatgccccttatgacctgggctacacacgtgctaca1200
atgggcagaacaaagggcagcgaagccgcgaggctaagccaatcccacaaatctgttctc1260
agttcggatcgcagtctgcaactcgactgcgtgaagctggaatcgctagtaatcgcggat1320
cagcatgccgcggtgaatacgttcccgggccttgtacacaccgcccgtcacaccacgaga1380
gtttgtaacacccgaagtcggtgaggtaaccttttggagccagccgccgaaggtgggaca1440
gatgattggggtgaagtcgtaacaagag1468
Claims (5)
1. Bacillus licheniformis, is characterized in that: be Bacillus licheniformis (Bacilluslicheniformis) ZJU-X1, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number is CGMCCNo.9688.
2. utilize Bacillus licheniformis as claimed in claim 1 to prepare and the Bacillus licheniformis preparation that obtains, it is characterized in that: form by as the Bacillus licheniformis ZJU-X1 of effective constituent and carrier;
Described carrier is at least one in maltodextrin, wheat bran, dregs of beans, stone flour, Semen Maydis powder;
Described Bacillus licheniformis preparation is as fodder additives.
3. utilize Bacillus licheniformis as claimed in claim 1 to prepare and the animal microorganism preparation that obtains, it is characterized in that: comprise the Bacillus licheniformis ZJU-X1 as activeconstituents I and carrier;
Described carrier is at least one in Semen Maydis powder, maltodextrin, wheat bran, dregs of beans, stone flour;
The microbiotic of described animal microorganism preparation alternatively in weaning pigs.
4. animal microorganism preparation according to claim 3, is characterized in that: also comprise active component II;
Described active component II is at least one in clostridium butylicum, subtilis, milk-acid bacteria, bifidus bacillus.
5. the animal microorganism preparation according to claim 3 or 4, is characterized in that:
In described animal microorganism preparation, the viable count of Bacillus licheniformis preparation is 10
11cfu/g.
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