CN106318890A - Phytase generating lactobacillus crispatus isolated from SPF (specific pathogen free) chicken craws and application thereof - Google Patents
Phytase generating lactobacillus crispatus isolated from SPF (specific pathogen free) chicken craws and application thereof Download PDFInfo
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Abstract
The invention provides phytase generating lactobacillus crispatus isolated from SPF (specific pathogen free) chicken craws and an application thereof. The collection number of lactobacillus crispatus MSY is CGMCC No.12928. The colony of the strain on an MRS plate is white. The strain has ragged edge, is flaky and has hemispherical heave in the middle. Through observation by gram staining, the strain is typically gram-positive, is more deeply stained, is microscopically shaped like a slender rod, has microbend appearing in a chain and does not have flagella or spores. The strain MSY obtained through isolation is a craw dominant colonized strain, has stronger acid resistance and certain bile salt tolerance and can be fermented in situ in the craws, secrete phytase and inhibit growth of escherichia coli and staphylococcus aureus. The strain is used for feeding animals and has the functions of increasing the absorptivity of calcium and phosphorus in feeds by animals, promoting animal growth and maintaining microecological balance in animal intestines.
Description
Technical field
The present invention relates to a strain Lactobacillus crispatus bacterial strain, specifically a strain is isolatable from the secretion of phytase of SPF chicken stomach
Lactobacillus crispatus bacterial strain and application.Belong to functional probiotic bacteria and applied technical field thereof.
Background technology
China's animal husbandry development is rapid in recent years, and abuse of antibiotics phenomenon is universal, and the abuse of antibiotics of edible animal is easy
Causing antibiotic to accumulate in animal body, develop immunity to drugs antibacterial, serious threat human health.At present, the country such as European Union is the tightest
Lattice limit even forbids the antibiotic use as growth promoter, promotes animal to use probiotic bacteria to solve these problems.Tradition meaning
The probiotic bacteria often function of justice general, does not has clear and definite and significant effect.
Phytic acid i.e. phytic acid, is the major storage form of phosphorus in the crop such as corn, beans, but nonruminant and people lack
The enzyme of weary decomposition phytic acid and be difficult by the phosphorus of this kind of form.In order to meet the needs of normal growth, metabolism, need to raise animal
Material adds Phos such as calcium hydrogen phosphate, not only causes the waste of phosphor resource, it is impossible to the follower feces digested and assimilated is discharged
External phytic acid easily causes environmental pollution；Meanwhile, phytic acid during animal gastrointestinal tract is digested and assimilated can with various metals from
Son is such as Zn2+、Ca2+、Cu2+、Fe2+And the trophic factors such as protein chelates into not capacitive complex, reduce the utilization of trophic factors
Rate, antinutritional factor of being known as.Owing to phosphorus ore resource is limited, relevant raw and auxiliary material appreciates and the raising of environmental requirement,
The production cost of calcium hydrogen phosphate is greatly improved.Phytase i.e. phytase, can be decomposed into inositol and Phos by phytic acid,
The utilization rate of phosphorus can be increased, reduce phosphorus and pollute and release the anti-oxidant action of phytic acid.
In the feedstuff of nonruminant, add the effect of phytase it is verified that, phytase becomes a kind of and replaces phosphoric acid hydrogen
The novel fodder additive of calcium, but phytase content in natural plants is relatively low, extraction cost is expensive；Build Pichia sp. work
Journey bacterial strain, fermenting and producing, extraction are transported and use also cost high, pollute and waste serious, and the biological characteristics of this enzyme is such as
Heat stability, protease inhibitor characteristic are low, it is impossible to the High Temperature High Pressure of the opposing pellet course of processing, have higher wanting to holding conditions
Ask.
The physiological function of birds crop is temporarily to store and soften food, is beneficial to birds and fully digests food and inhale
Receive.Crop is good endogenous probiotic bacteria habitat, surely grows a large amount of probiotic bacteria, and molecular ecology analysis shows Lactobacillaceae
Antibacterial accounts for 99.74% in crop mucosal sample, accounts for 62.75% and 3.09% in crop and cecal content respectively, and
Then accounting for 23.51% in feces, wherein Lactobacillus salivarius and Lactobacillus crispatus are the most universal, for maintaining animal intestinal micro-ecology to put down
Scale plays an important role.
If phytase generating probiotic bacteria can be separated to from crop, natural in crop surely grow, a large amount of, continue, send out in situ
Ferment, phytic acid of can directly degrading, discharge Phos, overcome the some shortcomings of vivoexpression phytase.As novel Tiny ecosystem
Additive, by the general layout of feed enzyme of fundamentally taking on a new look, cost-effective, reduce and pollute and waste, be the Gospel of feedstuff industry.
Summary of the invention
It is an object of the invention to solve deficiency of the prior art, it is provided that the curling breast bar of the secretion of phytase that a strain is new
Bacteria strain, this strains separation from specific pathogen free (being called for short SPF) chicken stomach, has the characteristic of phytase generating, may be used for developing
Exploitation animal probiotics preparation.
For achieving the above object, the present invention is by the following technical solutions:
Lactobacillus crispatus bacterial strain of the present invention, is isolatable from SPF chicken stomach content, streak culture through MRS solid medium, point
From obtaining strain Lactobacillus crispatus bacterial strain (Lactobacillus crispatus) MSY after purification, in JIUYUE in 2016 preservation on the 2nd
In China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), deposit number is CGMCC No.12928.
Present invention isolated above-mentioned Lactobacillus crispatus bacterial strain from SPF chicken stomach content by the following method: by nothing
The crop content taking from SPF chicken of bacterium distilled water diluting is rule on MRS solid medium, and the obvious bacterium colony of picking feature is anti-
Rule again separation, until isolating form, bacterium colony of the same size, carry out Gram’s staining, thalli morphology is carried out micro-sight
Examine.Repeatedly inoculate and obtain after purification.
This bacterial strain bacterium colony on MRS flat board is white, and edge is irregular, lamellar, center projections hemispherical, and gram contaminates
Color is observed, and this bacterial strain is typical case's Gram-positive, and dyeing is relatively deep, and in elongated rod-shaped under microscope, micro-bend chaining occurs, without whip
Hair, without spore.Well-grown in MRS fluid medium, in uniform turbid growth, occurs that white is heavy for 12 hours bottom test tube
Form sediment.Optimum growth temperature is 37 DEG C.
Through 16s rDNA sequence analysis, strain is identified, the Lactobacillus crispatus bacterial strain of institute of the present invention isolated
The classification position of MSY belongs to: Lactobacillus Lactobacillus crispatus (Lactobacillus crispatus).
The present invention separates the MSY bacterial strain of acquisition can be with secretion of phytase, and phytase activity is about 0.21U/ml.Can suppress
Escherichia coli and the growth of staphylococcus aureus, show that bacterial strain MSY of the present invention may be used for preparing antibacterial.This bacterial strain is at pH
The culture fluid of=3.0 still has certain growth, there is stronger capacity antacid, also there is certain bile tolerance ability.MSY bacterium
Agent feeding animals, can improve calcium phosphorus utilization, improves production performance, intestinal microecology balance.
There is advantages that
(1) MSY bacterial strain of the present invention, is accredited as Lactobacillus crispatus (Lactobacillus through 16s rDNA sequence analysis
Crispatus), this bacterial strain can secretion of phytase, phytase activity is about 0.21U/ml.Can divide for isolated first
Secrete the Lactobacillus crispatus of phytase.
(2) MSY bacterial strain of the present invention is that crop advantage grows bacterial strain surely, has acidproof, bile tolerance characteristic.By Odontothrips loti body
Outer bacteriostatic test shows, this bacterial strain is respectively provided with stronger antibacterial work to gram negative bacteria escherichia coli and gram positive bacteria
With, presenting broad-spectrum antimicrobial effect, this bacterial strain may be used for developing animal probiotics preparation.
(3) MSY bacterial strain of the present invention may be used for regulate microbial population of animal intestinal tract, by this microbial inoculum feeding animals (include chicken, duck,
The nonruminants such as pig) there is promotion growth, microecological balance in animal intestinal can be kept.Improve animal to Calcium in Feed
The absorbance of phosphorus, reduces the calcium phosphorus addition in daily ration and reduces calcium phosphorus emission in animal wastes, preserving the ecological environment.
(4) MSY bacterial strain of the present invention and the engineered strain of phytase generating and compared with feeding bacillus cereus, as feed additive
Feeding chickling has fine advantage: itself be that crop advantage grows bacterial strain, in-situ fermentation surely, it is not necessary to the various costs of factory；One
Secondary feed surely grow after, can breed continually and secretion of phytase, it is not necessary to constantly from feedstuff add；Inherent advantage bacterium
Strain, safe ready.
Phytase, as a kind of excellent food and feed additive, is more and more paid attention to.Due to natural origin
Phytase or extract difficulty, or secretory volume is the lowest, or cost is the highest, it is difficult to meets and produces needs.The most general logical
Cross and build the bacterial strain of energy efficiently Expressing Recombinant Phytase such as, pichia yeast phytase engineering bacteria phytase generating in next life.But in profit
There is also cost during carrying out fermenting and producing with this engineering bacteria, extract transport and use high, pollute and waste serious problem.And
And the biological characteristics of this enzyme such as heat stability, protease inhibitor characteristic are low, it is impossible to the high temperature of the opposing pellet course of processing is high
Pressure, has higher requirements to holding conditions.And the method feeding the bacillus cereus of phytase generating, equally exist needs the most past
Feedstuff adds the problem that bacillus cereus could meet animal needs.Thus, by comparing it can be seen that MSY bacterial strain of the present invention
There is obvious advantage.
Accompanying drawing explanation
Fig. 1 is Lactobacillus crispatus of the present invention (Lactobacillus crispatus) MSY Gram’s staining figure.
Fig. 2 is Lactobacillus crispatus of the present invention (Lactobacillus crispatus) MSY anti-microbial pathogen result of the test；
Wherein, A is that MSY suppresses escherichia coli (Escherichia coli ATCC 25922) growth result；
B is that MSY suppresses staphylococcus aureus (Staphylococcus aureus ATCC 25913) growth result.
Fig. 3 is Lactobacillus crispatus of the present invention (Lactobacillus crispatus) MSY acid resistance test result；
Fig. 4 is the 16s rDNA gene V3 district of Lactobacillus crispatus of the present invention (Lactobacillus crispatus) MSY,
Pcr amplification product electrophoretogram.
Detailed description of the invention
Below in conjunction with concrete test method and accompanying drawing, technical scheme and produced technique effect thereof are carried out
Further elucidated above, the description below is merely to explain the present invention, but is any limitation as the present invention never in any form, based on this
Any conversion of invention training centre work or replacement, belong to protection scope of the present invention.
Method used in the present invention if no special instructions, is this area conventional method.Used by following embodiment
Test material, reagent etc., if no special instructions, the most commercially obtain.
The separation of lactic acid bacteria in embodiment 1:SPF chicken stomach
Test material:
SPF chicken: specific pathogen free chicken (SPF), Shandong Province of China Shanxi Academy of Agricultural Sciences poultry SPF research center.
The preparation of culture medium:
MRS solid medium: for a kind of selective medium cultivated used by lactic acid bacteria, every L 10.0g Han peptone, cattle
Meat powder 5.0g, yeast powder 4.0g, glucose 20.0g, magnesium sulfate 0.1g, sodium acetate 5.0g, Triammonium citrate 2.0g, phosphoric acid hydrogen two
Potassium 2.0g, manganese sulfate 0.05g, tween 1.0g, agar 15g.
Test method:
(1) collection of sample
Slaughter 120 age in days SPF chickens, the aseptic crop opening chicken, take chicken stomach content with aseptic operation cutter and put into aseptic
Centrifuge tube in, dilute with physiological saline solution.
(2) separation and Culture of lactic acid bacteria in chicken stomach
From physiological saline solution diluent, take 3～4 inoculating loops, line on MRS solid selection medium respectively, then
Being placed in 37 DEG C of candle cylinders and cultivate after 12～24h, the obvious bacterium colony of picking feature is rule separation repeatedly respectively, until isolating shape
State, bacterium colony of the same size, carry out Gram’s staining by institute's isolated strains, thalli morphology carried out microexamination.Obtained strains
Named MSY.
Result of the test: the bacterium colony on MRS flat board is white, and edge is irregular, lamellar, center projections hemispherical, leather orchid
Albert'stain Albert is observed, and this bacterial strain is typical case's Gram-positive, and dyeing is relatively deep, and in elongated rod-shaped under microscope, micro-bend chaining occurs, nothing
Flagellum, without spore, result is shown in Fig. 1.
Embodiment 2: the screening of phytase-producing strain
Test material:
Sodium phytate: purchased from Sigma company.
Culture medium is prepared:
MRS fluid medium: MRS fluid medium is a kind of culture medium cultivated used by lactic acid bacteria, every L contains peptone
10.0g, beef powder 10.0g, yeast powder 5.0g, glucose 20.0g, magnesium sulfate 0.1g, sodium acetate 5.0g, ammonium citrate 2.0g,
Dipotassium hydrogen phosphate 2.0g, manganese sulfate 0.05g, tween 1.0g.
Test method:
(1) strain culturing
The bacterial strain of embodiment 1 isolated is inoculated in MRS liquid by the inoculum concentration of the 10% of MRS fluid medium volume
In culture medium, cultivate 48h for 37 DEG C.
(2) phytase activity of exocytosis is measured
Using with Spectrophotometric Methods For Determination of Phytic Acid enzymatic activity, its ultimate principle is that phytase is in uniform temperature and pH condition
Under, hydrolysis substrate sodium phytate, generate orthophosphoric acid and inositol derivative, in an acidic solution, at vanadium ammonium molybdate, comprehend generation Huang
[(NH4) of color3PO4NH4VO3·16MoO3] complex, under wavelength 415nm, carry out colorimetric determination.
Collect MSY bacterial strain bacterium solution, centrifuging and taking 200 μ L of supernatant, with 2mL phytase specificity substrate sodium phytate (pH5.0,
Sodium acetate buffer is prepared, final concentration of 5.0mM) mixing, after 37 DEG C of temperature bath 30min, terminate with 2mL vanadium ammonium molybdate stop buffer
Reaction, measures light absorption value at 415nm, measures extracellular phytase enzyme and lives.Sample sodium phytate concentration be 5.0mM, temperature be 37
DEG C, under conditions of pH is 5.0, per minute from sodium phytate, discharge 1 μm ol Phos, be a phytase activity unit, with U
Represent.
Result of the test: embodiment 1 separates the MSY bacterial strain obtained can be with secretion of phytase, and phytase activity is about 0.21U/
ml。
The biochemical identification of embodiment 3:MSY bacterial strain
Test material:
Indicator bacteria: escherichia coli (Escherichia coli ATCC 25922) and staphylococcus aureus
(Staphylococcus aureus 25913)。
Fel Sus domestica salt reagent: purchased from Beijing extensive and profound in meaning star Bioisystech Co., Ltd.
Culture medium is prepared:
Meat soup solid medium: every L 5g Han Carnis Bovis seu Bubali cream, peptone 10g, sodium chloride 5g, agar 20g.
Test method:
(1) the anti-microbial pathogen test of MSY bacterial strain:
Indicator bacteria bacteria suspension: take bacterium escherichia coli to be instructed (Escherichia coli ATCC 25922) and golden yellow
Staphylococcus (Staphylococcus aureus ATCC 25913) preserves liquid and carries out ruling (making on meat soup solid medium
It occurs that single bacterium colony is as the criterion), it is put in 37 DEG C of constant temperature culture 16h, the mono-bacterium colony of picking 0.8mm-1.0mm is in 1ml physiological saline solution
In, vibrate 1-2min by vortex oscillator, bacterial concentration now about 107CFU/ml。
The preparation of detection flat board: broth bouillon is added agar and is uniformly dissolved, autoclaving, it is cooled to about 50 DEG C,
Accurately measure this culture medium of 20ml and add in consistent and horizontal positioned in advance the culture dish of internal diameter, after solidification and make water in culture dish
Part is dried thoroughly.
The coating of indicator bacteria bacteria suspension: take indicator bacteria escherichia coli (the Escherichia coli of 20 μ l fresh cultured
ATCC 25922) and staphylococcus aureus (Staphylococcus aureus ATCC 25913) bacteria suspension (107CFU/
Ml) even spread on flat board, and with tweezers, aseptic Oxford cup is put in culture dish, gently in the plate of horizontal positioned
Place Oxford cup equably.
4. MSY bacteria suspension is added: in the cup of Oxford, be separately added into 80 μ l isolated strains MSY culture fluid, another Oxford cup
Middle MRS fluid medium aseptic for addition 80 μ l compares, and cultivates 24h for 37 DEG C.
Result of the test: isolated strains MSY can suppress the growth of escherichia coli and staphylococcus aureus, result is shown in Fig. 2.
Show that bacterial strain MSY of the present invention may be used for preparing antibacterial.
(2) acid resistance test of bacterial strain: connect separating MSY bacterial strain respectively by the inoculum concentration of the 1% of MRS fluid medium volume
Plant in the MRS fluid medium that pH value is 2.0,3.0,4.0,5.0,6.0 and 6.5, cultivate 24h for 37 DEG C, measure OD600nmValue.
Result of the test: bacterial strain of the present invention still has certain growth in the culture fluid of pH=3.0, has stronger antiacid energy
Power, result is shown in Fig. 3.
(3) the bile tolerance test of bacterial strain: the inoculum concentration inoculation that MSY bacterial strain presses the 1% of MRS fluid medium volume will be separated
Be respectively 0.03% in Fel Sus domestica salt content, 0.1%, 0.2%, 0.3%, MRS culture medium in, after 37 DEG C are cultivated 24h, dilute 10
Times, take 0.1ml and be coated with MRS solid plate, carry out viable plate colony counting.Colony counting method calculates viable count.
Result of the test: bacterial strain of the present invention has certain bile tolerance capability result and is shown in Table 1.
Table 1 Lactobacillus crispatus MSY is in the survival condition of various biliary salinity
The Species estimation of the 16S rDNA of embodiment 4:MSY bacterial strain
Test material: PCR reagent is purchased from TaKaRa biotech firm；Bacterial genomes DNA extraction kit (
Bacterial DNAKit) it is purchased from Omega company；Primer synthesis and order-checking are completed by the raw work in Shanghai.
Test method: utilize test kit to extract bacterial genomes DNA, using genomic DNA as template, utilize antibacterial 16s
RDNA universal primer 27F (primer sequence: SEQ ID NO.2) and 1492R (primer sequence: SEQ ID NO.3), amplification length is
The 16s rDNA variable region V3 region sequence of 1500bp, pcr amplification product is by Shanghai raw work order-checking, and gained sequence is with NCBI's
Known array in GeneBank compares.
PCR amplification system is 50 μ L:
PCR reaction condition is: 94 DEG C of denaturations 5min, 94 DEG C of degeneration 45s, 57 DEG C of annealing 30s, 72 DEG C of extension 2min, 30
Individual circulation；Last 72 DEG C extend 10min eventually；4 DEG C of preservations.
Result of the test: through comparison, gained sequence and Lactobacillus crispatus homology more than 99%, the sequencing results table
Bright, bacterial strain MSY of the present invention is Lactobacillus crispatus.16s rDNA gel electrophoresis figure, is shown in Fig. 4；16s rDNA nucleotide such as SEQ ID
Shown in NO.1.
The feeding experiment of embodiment 5:MSY bacterial strain
Test material:
Randomly choose 3 hen houses in large-scale broiler field, each individually raise table hens 500, conventional raise.
Feeding six and broiler fodder, normal feedstuff formula is shown in Table 2, retains the inorganic calcium phosphorus content on basis.Matched group 1 feeds
Normal feedstuff, matched group 2 adds KDN phytase 600U/KG with normal feedstuff, and test group adds MSY bacterium solution by drinking-water
107Cfu/ml, feeds normal feedstuff.
Test method:
(1) production performance test: feeding cycle is 6 weeks, respectively feeds a defective material, free choice feeding and drinking-water；Every day observes
The mental status of chicken, feces situation, during 42 age in days, the dead chicken quantity of statistics, it is calculated as motility rate；Add up three groups of respective feedstuffs always to disappear
Consumption and TBW, calculate feed-weight ratio.
Result of the test: matched group 1,2 survival rate is 97.5%, 96.8%, and feed-weight ratio is 2.04:1,2.03:1；Test group
Survival rate is 97.3%, and feed-weight ratio is 2.03:1.Show that inoculating MSY affects aspect to broiler performance, can reach directly to add
Add the production effect of phytase, be shown in Table 3.
Table 2 normal feedstuff formula and trophic level, calcium phosphorus content
Trophic level
Table 3 meat chicken production performance
(2) alcium and phosphor metabolization test:
Gather excrement sample respectively at 7,14,21 ages in days, measure excrement discharge calcium content by EDTA complexometry；Measure with ammonium molybdate method
The content of phosphorus.
1. the decomposition of sample: weigh 2～5g samples (accurately to 0.0002g), on electric furnace, low-temperature carbonization is to smokeless.
Put it into high temperature furnace calcination 3h under (550 ± 20 DEG C) again.In the crucible fill ash add 1:3 hydrochloric acid solution 10mL and
Concentrated nitric acid few drops, carefully boils.This solution is proceeded to 100mL volumetric flask, and with the filter in hot distilled water washing crucible and funnel
Paper, after being cooled to room temperature, constant volume, shake up, for sample decomposed solution.
2. sample calcium measures: accurately point take 10～20mL sample decomposed solution (calcic 2～25mg) in 150mL triangular flask,
Distilled water 50mL, 10g/L starch solution 10mL, triethanolamine solution 2mL, ethylenediamine solution 1mL, often adding a kind of reagent to fill
Dividing and shake up, then add malachite indicator 1, shake up, dropping 200g/L potassium hydroxide solution is to the most colourless, and repeated hydrogenation potassium oxide is molten
Liquid 2mL, adds 0.1g oxammonium hydrochloride., after shaking up dissolving, adds calcium Huang indicator a little, and it is blackish green for making color, in black background
Under, it is titrated to green fluorescence with 0.01mol/L EDTA standard solution immediately and disappears, be titration end-point in aubergine.Try simultaneously
Agent blank assay.
3. sample phosphorus measures: accurately pipettes sample decomposed solution 1-10mL (phosphorus content 50-70 μ g) and in 50mL volumetric flask, adds
Vanadium ammonium molybdate colour reagent 10mL, with distilled water diluting to scale, shakes up, and places more than 10min.With blank as reference, use
10mL colorimetric pool is under 420nm wavelength, with the absorbance of spectrophotometric determination sample decomposed solution.Sample is checked in standard curve
The phosphorus content of decomposed solution.
Result of the test: test group is close with the matched group 2 adding phytase, and without significant difference, the discharge of calcium phosphorus is less；With right
According to organizing 1 significant difference.
The calcium phosphorus content (%) of table 4 broiler excrement discharge
(3) on intestinal microbial population impact test:
Gathering caecum respectively at 14 ages in days, cut caecum opening by sterile scissors, carefully extrusion content is to centrifuge tube,
It is carried out continuously 10 times with physiological saline solution and is diluted to 10-8, select OK range dilution factor, with table 5 culture medium culturing, bacterium colony
Counting.
Result of the test: three groups of overwhelming majority strain differences are notable, and test group aerobe, enterobacteria, enterococcus are planted with interpolation
Acid enzyme matched group is close, with matched group 1 significant difference；Lactobacillus, bacillus bifidus, bacteroid, total anaerobe content successively
For test group, matched group 2 (phytase), matched group 1 (blank), and significant difference.
Table 5 cecum flora measures culture medium
Table 6 MSY affects (lgcfu/g feces) to 14 Day-old Broiler Chickens cecum flora
To sum up result of study shows: inoculates a small amount of MSY and has reached to add the production effect of KDN phytase 600U/KG, to meat
Chicken production performance is obviously improved；And the discharge of calcium phosphorus is less, improves the absorption rate of Calcium in Feed phosphorus；Improve in intestinal prebiotic
Bacterium: lactobacillus, bacillus bifidus, bacteroid, the content of total anaerobe.Can be seen that MSY bacterial strain of the present invention have include but not
It is limited to the purposes of following aspect: MSY microbial inoculum feeds chicken, duck, pig, calcium phosphorus utilization can be improved, improve production performance, the micro-life of intestinal
State balances.
Claims (7)
1. a strain Lactobacillus crispatus bacterial strain MSY, deposit number is CGMCC No.12928.
2. Lactobacillus crispatus bacterial strain MSY as claimed in claim 1, is characterized in that having the 16S shown in SEQ ID NO.1
RDNA sequence.
3. Lactobacillus crispatus bacterial strain MSY as claimed in claim 1 or 2, is characterized in that, described bacterial strain can secretion of phytase.
4. Lactobacillus crispatus bacterial strain MSY as claimed in claim 1 or 2, is characterized in that, described bacterial strain is that crop advantage grows bacterium surely
Strain.
5. the Lactobacillus crispatus bacterial strain MSY described in claim 1 reconciles microbial population of animal intestinal tract in preparation or promotes growth of animal
Application in additive.
6. the purposes in preparing probiotic bacteria of the Lactobacillus crispatus bacterial strain MSY described in claim 1.
7. the purposes described in claim 6, is characterized in that, described probiotic bacteria has anti-escherichia coli and staphylococcus aureus
Effect.
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| CN108546663A (en) * | 2018-05-10 | 2018-09-18 | 北京市农林科学院 | One boar source book song lactobacillus and its application |
| CN113652372A (en) * | 2021-08-30 | 2021-11-16 | 湖南师范大学 | Lactococcus lactis and application thereof in preparation of phytase and animal feed |
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| CN105518150A (en) * | 2013-09-06 | 2016-04-20 | 索发股份公司 | Method for evaluating effects of composition comprising microorganisms on intestinal microbiota |
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