CN104830806B - A kind of wide fragmentation pattern salmonella bacteriophage and its antibacterial application - Google Patents
A kind of wide fragmentation pattern salmonella bacteriophage and its antibacterial application Download PDFInfo
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Abstract
The present invention relates to one plant of bacteriophage and its antibacterial application, especially one plant wide fragmentation pattern salmonella bacteriophage and its antibacterial application, belong to bioengineering field.One plant wide fragmentation pattern salmonella bacteriophage, preserving number is:CCTCC NO：M 2014145, preservation date is on April 24th, 2014, and the bacteriophage has strong splitting action to salmonella.In the present invention, this plant of bacteriophage is analyzed from electron microscopic morphology, belongs to Myoviridae, is named as salmonella bacteriophage STP4 a；It is that can survive under conditions of 4 12 in 40 60 DEG C and pH；20 DEG C preserve 1 year after it is activity stabilized；The protective agent that bacteriophage preserves is the culture solution containing 20% glycerine.Salmonella with drug resistance can be killed using the phage specificities cracking salmonella of the present invention.
Description
Technical field
The present invention relates to one plant of bacteriophage and its antibacterial application, especially a kind of wide fragmentation pattern salmonella bacteriophage and its
Antibacterial application, belongs to bioengineering field.
Background technology
Salmonella (Salmonella) is Gram-negative straight-bar bacterium, is a kind of important infecting both domestic animals and human pathogen.State
Inside and outside research shows, is ranked forefront repeatly position by salmonellal food origin disease.With a large amount of of antibiotic and be used for a long time,
In food and environment, can the drug-resistant type salmonella that be on the rise have influenceed the treatment of human infection's salmonella disease, grind
It is the challenge that current scientific research personnel faces to study carefully new biological bacteriostatic agent to substitute chemobiotic.
After bacteriophage is a class bacterial virus, virulent phage bacterial infection, can rapid cleavage bacterium cause Host Strains
Death.Salmonella bacteriophage with specific infection and can crack salmonella, thus with bactericidal action.In infection model
In, bacteriophage specific can kill salmonella, and in food, bacteriophage can control bacterium dirty as antiseptic
Dye, with potential Development volue.
The content of the invention
The present invention provides a kind of wide fragmentation pattern salmonella bacteriophage, has the bacteriophage that fine melt is acted on to salmonella
Preparation, said preparation can individually or compounding use, can the specific a variety of salmonellas of inactivation, be current prevention and control salmonella
Pollution provides a kind of bacteriophage rule of origin safe and free of toxic and side effects.
The present invention also provides a kind of antibacterial application of the wide fragmentation pattern salmonella bacteriophage.
The technical solution adopted for the present invention to solve the technical problems is：
A kind of wide fragmentation pattern salmonella bacteriophage, preserving number is:CCTCC NO：M 2014145, preservation date is 2014
In on April 24, in, the bacteriophage has strong splitting action to salmonella.In the present invention, this plant of bacteriophage is from electron microscopic morphology
Analysis, belongs to Myoviridae, is named as salmonella bacteriophage STP4-a；In the bar that 40-60 DEG C and pH is 4-12
It can be survived under part；- 20 DEG C preserve 1 year after it is activity stabilized；The protective agent that bacteriophage preserves is molten for the culture containing 20% glycerine
Liquid.
A kind of application of bacteriophage in the medicine for killing salmonella is prepared, it is characterised in that：By salmonella
Bacteriophage after purification, for suppressing and killing salmonella.
The bacteriophage is used to prevent salmonella-polluted and salmonella undue growth, wherein described salmonella includes
Salmonella typhimurium, Salmonella paratyphi A, Bacterium enteritidis, salmonella typhi and B-mode Salmonella typhimurium
Bacterium.
A kind of application of described wide fragmentation pattern salmonella bacteriophage in terms of food pollution salmonella is prevented and treated.
After bacteriophage is diluted with water, as flushing liquor or leacheate, for the production environment to food or production utensil
Or production equipment carries out sprinkling dissipation, for controlling pollution of the salmonella to described environment or utensil or equipment.
The food refers to：Selected in grain, meat, vegetables, eggs one fabricated product, or by they combine processing
Product.
A kind of phage preparation containing described wide fragmentation pattern salmonella bacteriophage.The phage preparation is to Salmonella
Bacterium has a fine melt effect, said preparation can individually or compounding use, can the specific a variety of salmonellas of inactivation, be current
Prevention and control are salmonella-polluted to provide a kind of bacteriophage rule of origin safe and free of toxic and side effects.
When bacteriophage is with 109The addition of pfu/g feeds is added in chicken feed, can make the number of salmonella in chicken intestinal
Amount 1 order of magnitude of reduction, fungistatic effect significantly, does not influence then on other bacteriums.
The beneficial effects are mainly as follows：It can be killed with resistance to using phage specificities cracking salmonella
The salmonella of the property of medicine.
Brief description of the drawings
Fig. 1 is bacteriophage electromicroscopic photograph (× 40K)；
Fig. 2 is bacteriophage temperature sensitivity testing result；
Fig. 3 is bacteriophage pH sensitivity Detection results；
Fig. 4 is bacteriophage storage temperature analysis chart；
Fig. 5 is separate groups of mice liver tissue slices figure (H.E × 400)；
Fig. 6 is separate groups of mice spleen tissue slice map (H.E × 200)；
Fig. 7 is separate groups of mice renal tissue slice map (H.E × 400)；
Fig. 8 is separate groups of mice gastric tissue slice map (H.E × 200)；
Fig. 9 is separate groups of mice intestinal tissue slice map (H.E × 200)；
Figure 10 is separate groups of mice heart tissue sections figure (H.E × 200)；
Figure 11 is different disposal group, salmonella content in different feeding time chicken intestinal.
Embodiment
Below by specific embodiment, technical scheme is described in further detail.It should be appreciated that this hair
Bright implementation is not limited to the following examples, and any formal accommodation and/or change made to the present invention will all fall
Enter the scope of the present invention.
In the present invention, if not refering in particular to, all part, percentage are unit of weight, equipment and raw material for being used etc.
It is commercially available or commonly used in the art.Method in following embodiments, is the normal of this area unless otherwise instructed
Rule method.
The bacterial strain that present invention experiment is related to：Two plants of salmonella typhimuriums (preserving number be respectively ATCC14028 and
ATCC13311), Salmonella paratyphi A (preserving number be CMCC (B) 50001), (preserving number is Bacterium enteritidis
CMCC50041), salmonella typhi (preserving number is CMCC50071) and B-mode salmonella typhi (preserving number is
CMCC50094 U.S. ATCC centers and China General Microbiological culture presevation administrative center) are purchased from respectively.
This bacteriophage STP4-a is stored in China typical culture collection center, address：Wuhan, China, Wuhan University, protect
Hide numbering：CCTCC NO：M 2014145, taxology name：Salmonella bacteriophage STP4-a (Salmonella
bacteriophage STP4-a)。
Embodiment 1：Bacteriophage separation is prepared and purified
Salmonella typhimurium ATCC14028 is seeded in 10mL nutrient broths, overnight shaking culture obtains Host Strains
Liquid.Next day take the sewage sample of Qingdao of Shandong province different location respectively with after host's bacterium solution mixed culture, centrifugation (3318g,
10min) take supernatant to obtain the solution containing bacteriophage by miillpore filter (0.22 μm) is degerming, take solution 0.1mL, carry out
10 times of dilutions, take each 0.1mL of dilution and host's bacterium solution 0.1mL of incubated overnight to mix, and add about 5mL semisolid culturemediums,
Nutrient agar panel upper strata is poured into rapidly after mixing, horizontal 5min is shaken up, treats that it solidifies, is placed in 37 DEG C of constant incubator cultures
Observed after 12h, obtain the double-layer plate for forming plaque, then continuously repeated at least 3 times single spots and puncture the pure of progress bacteriophage
Change, bacteriophage after purification is added in host's bacterium solution and carries out Multiplying culture, centrifugation (7607g, 20min) supernatant is through micropore
Bacteriophage stoste is obtained after filter membrane (0.22 μm) is degerming.
Semisolid culturemedium：By 19g/L nutrient broth mediums and 33g/L nutrient agars according to 1:1 volume
Than being formulated.
Bacteriophage electron microscopic observation
By prophage drop in paraffin plate, copper sheet net is placed in dropping liquid, many extraction raffinates are sucked after 15min with filter paper
Body, sucks surplus liquid after dripping phosphotungstic acid aqueous solution, dyeing 10min, 10-15min is dried in atmosphere, in transmission electron microscope
The form of lower observation bacteriophage.As shown in figure 1, bacteriophage STP4-a is tadpole shape, head length is 100nm, and transverse diameter is 80nm,
The elongate tail for being about 120nm with length, belongs to Myoviridae.
The influence of the temperature of embodiment 2 and pH to bacteriophage
EP pipes containing 900 μ L fluid nutrient mediums are put in the water-bath of different temperatures value and preheat 60min, temperature stabilization is treated
Afterwards, 100 μ L bacteriophage stostes are added, phage titer is determined after 60min is cultivated；By the liquid training of the different pH value of 900 μ L
Foster base is added separately in 2mL EP pipes, and it is to preheat 30min in 37 DEG C of constant incubator to be placed in temperature, after after temperature stabilization,
100 μ L bacteriophage stostes are added, phage titer is determined after 60min is cultivated.
As a result as shown in Fig. 2 the activity of bacteriophage keeps constant in 30min under the conditions of 40 DEG C；Under the conditions of 50 DEG C
Still there is 90% and 75% activity during 15min and 30min respectively, but pass through 60-80 DEG C, after 15min processing, the activity of bacteriophage
Only remain less than 0.1%.After 30min processing, the survival number of bacteriophage is less than 10pfu/mL.
As a result as shown in figure 3, STP4-a is under conditions of pH is 4-7, survival rate is 100%.When pH is 8-10, bacteriophage
Survival rate still have 90%.When pH is 11 and 12, the survival rate of bacteriophage is respectively 70% and 56%.
The bacteriophage preservation condition of embodiment 3 is analyzed
The glycerine that 1. bacteriophage adds purified phage solution final concentration of 20% is preserved using different methods
After be stored under the conditions of 4 DEG C and -20 DEG C.Stood after the skim milk that 2. purified phage solution is added to final concentration of 30%
It is i.e. lyophilized with liquid nitrogen, it is stored under the conditions of -80 DEG C.Judge preservation side by determining the potency of bacteriophage after preserving 1 year respectively
The feasibility of method.
As a result as shown in figure 4, -20 DEG C of Cord blood effects are best.4 DEG C after 1 year, are bitten with -80 DEG C of Cord blood bacteriophages
The potency of thalline reduces nearly 2 and 1 order of magnitude respectively.
The phage splitting analysis of spectrum of embodiment 4
By microbionation to 10mL fluid nutrient mediums, shaken cultivation to logarithmic phase respectively takes 100 μ L bacterium solutions to be added to 5mL half
In solid medium, it is poured onto in the flat board containing one layer of culture medium and forms double-layer plate, after dry, by the 10 of 10 μ L2Extremely
1010The bacteriophage point of pfu/mL potency is dropped on flat board, is inverted culture to plaque for 37 DEG C and occurs.
As a result as shown in table 1, using Host Strains salmonella typhimurium ATCC14028 as reference, STP4-a can be with contour
Effect ground cracks the salmonella of remaining 5 plants of different serotypes, and the phage solution of different diluted concentrations is all containing salmonella
The high plaque of Clear ＆ Transparent degree is formd on flat board, display STP4-a has stronger cracking to the salmonella of different serotypes
Effect.
The phage splitting analysis of spectrum of table 1
Remarks：+ represent generation transparent circle, i.e., with cracking ability ,-representative does not produce transparent circle, i.e., without cracking energy
Power.
The salmonella bacteriophage STP4-a of embodiment 5 Oral toxicity experiment
BALB/c mouse Beijing Vital River Experimental Animals Technology Co., Ltd..
Nutrient agar, Beijing Luqiao Technology Co., Ltd.；
Nutrient broth medium, Beijing Luqiao Technology Co., Ltd..
Tanon-4200SF fully automatic digital gel chemistries luminescence image analysis systems, Guangzhou reputation dimension biotechnology instrument has
Limit company.
UNIQ-10 pillar bacterial genomes DNA extraction agent boxes, Shanghai Sheng Gong bioengineering limited company；
DNase, Beijing Suo Laibao Science and Technology Ltd；
RNase, Beijing Suo Laibao Science and Technology Ltd；
Bar-shaped feed, Qingdao City institute for drug control.
Main agents are prepared
SM buffer solutions：Accurately weigh 5.8g NaCl, 2g MgSO4·7H2O is dissolved in 800mL distilled waters, plus 50mL
1mol/L Tris-HCl (pH 7.5), plus distilled water is to 1000mL, 121 DEG C of autoclaving 15min, room temperature preservation.
1mol/L Tris-HCl (pH 7.5) solution：Accurate 121.1g Tris plus the distilled water of weighing adjusts pH to 800mL
To 7.5, addition distilled water to 1000mL, 121 DEG C of autoclaving 15min, room temperature preservation.
Salmonella bacteriophage STP4-a Oral toxicity experiment
Phage propagation to potency is 1.5 × 1010Pfu/mL (solvent is SM buffer solutions), is placed in serum bottle in 4 DEG C of ice
Saved backup in case；
BALB/c mouse totally 30 (male mouse 15, female mice 15) is bought, 6-8 week old, body weight 16-22g is put in Animal House
Endoadaptation environment one week.After environment is adapted to, every mouse is weighed before gavage, and is marked with 3-5% picric acid.This reality
Test and corresponding disposable gavage processing is carried out to mouse, processing packet is shown in Table 2, and mouse needs fasting to can't help water 3-4h before contamination, prohibits
Need to weigh to animal before contaminating after food, still need to continue fasting 1-2h after contamination.In feeding process, the room temperature control of Animal House
At 22 ± 3 DEG C, relative humidity is 30-70%.Artificial light source (bright+10h of 14h are dark), animal edible standard are preferably used in receptacle
Feed, free water.
Processing mode of the different grouping of table 2 to mouse
Note：F, M represent female mice group, male mouse group respectively；1st, 2,3 blank group, negative control group, bacteriophage processing are represented respectively
Group.
30min and 4h that should be after contamination on the day of to animal contaminated respectively do once careful clinical observation, after
Once a day, Continuous Observation 14d.
(1) body weight：Should be weighed every animal after contamination, and body weight determination is carried out every other day, and carry out body weight record, calculate body
Change again；
(2) gross examination of skeletal muscle：Observation should include skin, hair, eyes and mucous membrane change, should also observe breathing, circulation, plant
The sign of nerve and central nervous system, limb activity situation and Behavioral change；
(3) histopathologic examination：All contamination animal (including dead or execution animal during experiment) is carried out for reply
Gross anatomy, records the weight of liver, kidney, spleen and thymus gland, calculates organ coefficient (organ coefficient=organ weights/body
Weight × 100%)；The macroscopic disease damage organs of record whole (observation organ includes liver, kidney, spleen, stomach, intestines, heart,
Preserved in 40% formalin), tissue specimen is through drawing materials, fixing, be dehydrated transparent, waxdip, FFPE, section, bush
Essence-Yihong (H.E) decoration method, mounting (in the strong Tilapia Fillets ozone sterilization treatments of Zhao Yong the generation of free radical and its to production
The influence of product quality and security:[Ph.D. Dissertation] Shandong：Food Science and Engineering institute of Chinese Marine University,
2013.), and histopathologic examination is done.
The group difference of data analysing method two is examined to be examined using t-, P<0.01 is judged as with pole significant difference, P<
0.05 is judged as with significant difference, P>0.05 is that difference is not notable.Test data using average value ± standard deviation ()
Represent.
Experimental result
Each 10 mouse of blank group, negative control group, experimental group are no dead existing during gavage and in 14 day observation period
As and each group mouse skin, hair, eyes and mucous membrane no abnormality seen, breathing, circulation, vegetative nerve and central nervous system
Also phenomenon without exception occurs for sign, limb activity situation and Behavioral change.Table 3, table 4, table 5 represent respectively each group mouse weight,
The statistical result of internal organs weight and organ coefficient.
(1) changes of weight
Bacteriophage group, bacteriophage group mouse weight are inactivated in each minute point compared with SM buffer control groups, through t-
Check analysis all there was no significant difference (P>0.05), illustrate that bacteriophage and its residue do not make significant difference to mouse weight, as a result
It is shown in Table 3.
The each group mouse weight statistical result of table 3 (g,)
Note：Subscript represents letter before significant difference result, ", " and represents F2 and F1 and M2 and M1 comparative results, ", " in table
Letter F3 and F1 and M3 and M1 comparative results afterwards, same letter represent that there was no significant difference (P ＞ 0.05), and different letters represent tool
There is significant difference (P ＜ 0.05), "-" represents to have neither part nor lot in comparison procedure.
(2) organ weights
Bacteriophage group, inactivation bacteriophage group are compared with SM buffer control groups, through t- check analyses, kidney, spleen, thymus gland
There was no significant difference for weight (P ＞ 0.05), illustrates bacteriophage and its residue to mouse kidney, spleen, thymic weight without notable
Influence.Wherein, t- check analyses are found, F2 group liver weights are significantly higher than F1 groups (P ＜ 0.05), illustrate the residue of bacteriophage
Liver growth will not be had a negative impact, it is possible to liver development can be promoted.It the results are shown in Table 4.
The each group mice organs of table 4 weight result (g,)
Note：Subscript represents letter before significant difference result, ", " and represents F2 and F1 and M2 and M1 comparative results, ", " in table
Letter F3 and F1 and M3 and M1 comparative results afterwards, same letter represent that there was no significant difference (P ＞ 0.05), and different letters represent tool
There is significant difference (P ＜ 0.05), "-" represents to have neither part nor lot in comparison procedure.
(3) organ coefficient
Bacteriophage group, inactivation bacteriophage group are compared with SM buffer control groups, through t- check analyses, kidney, spleen, thymus gland
There was no significant difference for coefficient (P ＞ 0.05), illustrates bacteriophage and its residue to mouse kidney, spleen, thymus gland coefficient without notable
Influence.Wherein, t- check analyses are found, F2 group liver weights are significantly higher than F1 groups (P ＜ 0.05), illustrate the residue of bacteriophage
Liver growth will not be had a negative impact, it is possible to liver development can be promoted.It the results are shown in Table 5.
The each group mice organs coefficient results of table 5 (%,)
Note：Subscript represents letter before significant difference result, ", " and represents F2 and F1 and M2 and M1 comparative results, ", " in table
Letter F3 and F1 and M3 and M1 comparative results afterwards, same letter represent that there was no significant difference (P ＞ 0.05), and different letters represent tool
There is significant difference (P ＜ 0.05), "-" represents to have neither part nor lot in comparison procedure.
(4) pathological examination result
A. gross anatomy result
Internal organs face is not found after all mouse dissection of blank group, inactivation bacteriophage treatment group and bacteriophage treatment group
There is anomaly in color, structure etc..Then, one mouse of every group of random picking make tissue pathological slice further look at whether
There is anomaly.
B. histopathological examination result
F1, F2, F3 group section between and M1, M2, M3 group cut into slices between no significant difference, Fig. 5, Fig. 6, Fig. 7, Fig. 8, figure
9th, Figure 10 represents the histotomy figure of different groups of liver, spleen, kidney, stomach, intestines and heart respectively.Wherein liver organization knot
Structure is normal, and no extravasated blood, ischemic, oedema, necrosis, fibrosis, envelope are complete.Lobuli hepatis, central vein is located at centrilobular, tube wall
Completely, there is hepatic sinusoid remittance.Liver rope, liver cell monolayer alignment is constituted, and is shown greatly centered on central vein to surrounding into radial
Arrangement；Spleen tissue structure is normal, no extravasated blood, oedema, hyperplasia；Renal tissue structure is normal, no oedema, necrosis, fibrosis.Skin
Visible glomerulus in matter, glomerulus structure is normal, and number is normal, and visible a large amount of different tangent planes, intensive arranged in parallel in medullary substance
Renal tubule, no Malpighian corpuscle；Have no that bleeding, necrosis, erosion or ulcer are formed in gastric tissue；The arrangement of intestinal tissue submucosal glands is still whole
Together, muscle layer and outer membrane structure are complete；Heart tissue structure is normal, no necrosis, fibrosis.Cardiac muscle fibre takes on a red color, cardiac muscle fibre
Between visible a small amount of loose connective tissue and a large amount of capillaries.Identical knot can also be drawn by being respectively organized in the section of M1, M2, M3 group
By.From histopathologic slide's result, each group histotomy figure is without significant difference, it is seen that bacteriophage STP4-a is to each organ group
No obvious negative effect is knitted, this and Carlton (Carlton R M, Noordman W H, Biswas B, et
al.Bacteriophage P100for control of Listeria monocytogenes in foods:Genome
sequence,bioinformatic analyses,oral toxicity study,and application[J]
.Regulatory Toxicology and Pharmacology.2005,43(3):301-312.)、Kang(Kang H W,
Kim J W,Jung T S,et al.wksl3,a New biocontrol agent for Salmonella enterica
serovars enteritidis and typhimurium in foods:characterization,application,
sequence analysis,and oral acute toxicity study[J].Applied and Environmental
Microbiology.2013,79(6):1956-1968.)、Mccallin(Mccallin S,Alam S S,Barretto C,
et al.Safety analysis of a Russian phage cocktail:from metagenomic analysis
to oral application in healthy human subjects[J].Virology.2013,443(2):187-
196.) result of study is consistent, and the security of bacteriophage is all demonstrated in terms of zoopery.
The security for further illustrating bacteriophage STP4-a is tested in this experiment by Oral toxicity, and data analysis finds phagocytosis
Body treatment group, inactivation bacteriophage treatment group are compared with blank group is in terms of body weight, organ weights, internal organs development all without notable
Sex differernce.And do not find that mouse takes intracorporeal organ after bacteriophage and occurs any disease by gross anatomy and histotomy observation
Become phenomenon.From the point of view of molecular level analysis and Oral toxicity experimental result, bacteriophage does not have obvious damaging effect, is a kind of peace
Full antiseptic.
Embodiment 6：Bacteriostatic experiments of the salmonella bacteriophage STP4-a in laying hen body
36 17 age in days laying hens are divided into 3 groups, such as table 6.In addition to A groups, every chicken difference intragastrically mouthful of remaining each group
Take 107Cfu salmonella typhimuriums ATCC14028.Bacteriophage is mixed with feed, its concentration is 109Pfu/g feeds, from microbiological contamination
3 days feeding body feedstuffs containing phagocytosis, continuous again after microbiological contamination to feed 14 days before, at the 7th day and the 14th day per dead 6 of component other places
Chicken, takes out its enteron aisle to determine bacterium number.Due to containing the advantage gut floras such as Escherichia coli in chicken intestinal, it is contemplated that considerably beyond body
Salmonella ATCC14028 quantity is inside colonized, salmonella is first passed through when then determining and increases bacterium processing, is increased by determining after bacterium
Salmonella bacterium number situation of change understands whether bacteriophage STP4-a has remarkable effect effect in chicken intestinal.Assay method is such as
Under：1 (g) is pressed into small intestine:In 10 (mL) immersion magnesium chloride peacock green (RV) enrichment liquids, sterile homogeneous is after 42 DEG C of cultures
After 24h, sample is subjected to ten times of gradient dilutions with sterile saline, selects appropriate dilution to take 100 μ L to be applied to xylose and relies ammonia
On sour deoxidation cholate (XLD) flat board, salmonella colony counting is carried out after 37 DEG C of culture 24h.
The experimental design of table 6
The salmonella in chicken intestinal at the 7th day and the 14th day respectively to different disposal group counts, and A groups are all not
Salmonella is detected, thus the salmonella in B, C group enteron aisle is all oral salmonella.Different disposal group, different raisings
Salmonella content results are shown in Table 11 in time chicken intestinal, wherein " * * " show that bacteriophage treatment group and positive controls have and extremely shown
Write difference, P<0.01；Cube represents bacterium number in every chicken intestinal, and horizontal line represents the average of bacterium number in 6 chicken intestinals of the group.
As shown in Figure 11, at the 7th day, C group Salmonella bacterium numbers have dropped 0.25log (cfu/g) compared with B groups, be examined through t-
Analysis, P>0.05, decline effect not notable.During by 14 days, C group Salmonella bacterium numbers have dropped 1 order of magnitude compared with B groups,
Through t- check analyses, decline effect extremely significantly (P ＜ 0.01), this explanation bacteriophage can play in chicken body certain controls curative effect
Really.
Embodiment described above is a kind of preferably scheme of the present invention, not makees any formal to the present invention
Limitation, also has other variants and remodeling on the premise of without departing from the technical scheme described in claim.
Claims (6)
1. one plant wide fragmentation pattern salmonella bacteriophage, it is characterised in that：Preserving number is：CCTCC NO：M 2014145, this is bitten
Thalline has effective splitting action to salmonella.
2. a kind of application of bacteriophage as claimed in claim 1 in the medicine for killing salmonella is prepared, it is characterised in that：Will
Salmonella bacteriophage after purification, for suppressing and killing salmonella.
3. application according to claim 2, it is characterised in that：The bacteriophage is used to prevent salmonella-polluted and Salmonella
Bacterium undue growth, wherein described salmonella includes salmonella typhimurium, Salmonella paratyphi A, Salmonella
Bacterium, salmonella typhi and B-mode salmonella typhi.
4. a kind of the answering in terms of food pollution salmonella is prevented and treated of the wide fragmentation pattern salmonella bacteriophage described in claim 1
With.
5. application according to claim 4, it is characterised in that：After bacteriophage is diluted with water, flushing liquor or elution are used as
Liquid, carries out sprinkling dissipation, for controlling salmonella to institute for the production environment to food or production utensil or production equipment
The environment or utensil stated or the pollution of equipment.
6. the application according to claim 4 or 5, it is characterised in that：The food refers to：By grain, meat, vegetables, egg
Selected in class one fabricated product, or by they combine fabricated product.
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| CN105950568A (en) * | 2016-06-14 | 2016-09-21 | 苏州埃瑞特生物技术有限公司 | High-quality bacteriophage preservation protective agent and preparation method and application thereof |
| CN106497888B (en) * | 2016-10-29 | 2018-06-29 | 华中农业大学 | Salmonella bacteriophage and bacteriophage bactericidal composition and its application |
| CN108359644B (en) * | 2018-02-07 | 2019-08-02 | 青岛诺安百特生物技术有限公司 | A kind of wide range salmonella bacteriophage and its application |
| CN108588037B (en) * | 2018-04-20 | 2021-01-05 | 华中农业大学 | Salmonella bacteriophage LPSE34 and application thereof in food |
| CN108546685B (en) * | 2018-04-20 | 2021-01-05 | 华中农业大学 | Salmonella enteritidis bacteriophage LPSE28 and application thereof in food |
| KR102068591B1 (en) * | 2018-05-03 | 2020-01-21 | 경북대학교 산학협력단 | Novel recombinant protein from bacteriophage having antibacterial activity to pathogenic gram-negative bacterium |
| CN109825479B (en) * | 2019-02-28 | 2020-05-01 | 华中农业大学 | Wide-spectrum salmonella bacteriophage LPSTLL and application |
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