CN104830806A - Salmonella bacteriophage being wide in lysis spectrum and bacterial inhibition application thereof - Google Patents
Salmonella bacteriophage being wide in lysis spectrum and bacterial inhibition application thereof Download PDFInfo
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Abstract
The invention relates to a bacteriophage and a bacterial inhibition application thereof, and especially relates to a salmonella bacteriophage being wide in lysis spectrum and a bacterial inhibition application thereof and belongs to the field of biological engineering. The salmonella bacteriophage being wide in lysis spectrum is assigned the accession number CCTCC NO: M2014145 at 2014 Apr.24. The salmonella bacteriophage has a strong lysis effect on salmonella. In the invention, electron microscopic morphologic analysis of the salmonella bacteriophage proves that the salmonella bacteriophage belongs to the family of myoviridae and is named salmonella bacteriophage STP4-a. The salmonella bacteriophage can survive even at 40-60 DEG C and under the pH of 4-12, and is stable in activity after storage at -20 DEG C for one year. A protective agent of the salmonella bacteriophage is a culture solvent containing 20% of glycerin. The bacterial inhibition, through special lysis of the salmonella, can kill the salmonella having drug resistance.
Description
Technical field
The present invention relates to a strain phage and antibacterial application thereof, especially a kind of wide fragmentation pattern Salmonellas phage and antibacterial application thereof, belong to bioengineering field.
Background technology
Salmonellas (Salmonella) is Gram-negative straight-bar bacterium, is a kind of important infecting both domestic animals and human cause of disease bacterium.Domestic and international research shows, to be ranked forefront repeatly position by salmonellal food origin disease.Along with antibiotic a large amount of and life-time service, in food and environment, the drug-resistant type Salmonellas be on the rise has affected the treatment of human infection Salmonellas disease, and can working out novel biological bacteriostatic agent, to carry out alternative chemobiotic be the challenge that current scientific research personnel faces.
Phage is a bacterioid virus, after virulent phage bacterial infection, can cause the death of Host Strains by rapid cleavage bacterium.Salmonellas phage can specific infection cracking Salmonellas, thus has germicidal action.In infection model, phage specificity can kill Salmonellas, and in food, phage can control bacterial contamination as antiseptic-germicide, has potential Development volue.
Summary of the invention
The invention provides a kind of wide fragmentation pattern Salmonellas phage, Salmonellas is had to the phage preparation of fine melt effect, said preparation can independent or composite use, can the multiple Salmonellas of specific deactivation, a kind of phage rule of origin safely, to have no side effect is provided for current prevention and control are salmonella-polluted.
The present invention also provides the antibacterial application of a kind of described wide fragmentation pattern Salmonellas phage.
The technical solution adopted for the present invention to solve the technical problems is:
A kind of wide fragmentation pattern Salmonellas phage, preserving number is: CCTCC NO:M 2014145, and preservation date is on April 24th, 2014, and this phage has strong splitting action to Salmonellas.In the present invention, this strain phage is analyzed from electron microscopic morphology, belongs to Myoviridae, by its called after Salmonellas phage STP4-a; Can both survive under 40-60 DEG C and pH are the condition of 4-12; Preserve after 1 year activity stabilized for-20 DEG C; The protective material that phage preserves is the culture solution containing 20% glycerine.
Described phage, preparing the application killed in the medicine of Salmonellas, is characterized in that: after Salmonellas phage purifying, for suppressing and killing Salmonellas.
This phage is used for preventing salmonella-polluted and Salmonellas hypertrophy, and wherein said Salmonellas comprises Salmonella typhimurium, Salmonella paratyphi A, Salmonella enteritidis, salmonella typhi and B-mode salmonella typhi.
The application of a kind of described wide fragmentation pattern Salmonellas phage in control food contamination Salmonellas.
After phage dilute with water, as flushing liquor or leacheate, for the production environment of food or produce utensil or production unit carries out sprinkling dissipation, for controlling the pollution of Salmonellas to described environment or utensil or equipment.
Described food refers to: by the fabricated product selecting in grain, meat, vegetables, eggs, or the fabricated product combined by their.
A kind of phage preparation containing described wide fragmentation pattern Salmonellas phage.This phage preparation has fine melt effect to Salmonellas, and said preparation can independent or composite use, can the multiple Salmonellas of specific deactivation, provides a kind of phage rule of origin safely, to have no side effect for current prevention and control are salmonella-polluted.
When phage is with 10
9the addition of pfu/g feed adds in chicken feed, and the quantity of Salmonellas in chicken intestinal can be made to reduce by 1 order of magnitude, and fungistatic effect is remarkable, does not then affect other bacteriums.
Beneficial effect of the present invention is mainly reflected in: adopt phage specificities cracking Salmonellas can kill the Salmonellas with resistance.
Accompanying drawing explanation
Fig. 1 is phage electromicroscopic photograph (× 40K);
Fig. 2 is phage temperature sensitivity detected result;
Fig. 3 is phage pH sensitivity Detection result;
Fig. 4 is phage 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 stomach-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 Salmonellas content in different treatment group, different feeding time chicken intestinal.
Embodiment
Below by specific embodiment, technical scheme of the present invention is described in further detail.Should be appreciated that enforcement of the present invention is not limited to the following examples, any pro forma accommodation make the present invention and/or change all will fall into scope.
In the present invention, if not refer in particular to, all parts, per-cent are weight unit, and the equipment adopted and raw material etc. all can be buied from market or this area is conventional.Method in following embodiment, if no special instructions, is the ordinary method of this area.
The present invention tests the bacterial strain related to: two strain Salmonella typhimuriums (preserving number is respectively ATCC14028 and ATCC13311), Salmonella paratyphi A (preserving number is CMCC (B) 50001), Salmonella enteritidis (preserving number is CMCC50041), salmonella typhi (preserving number is CMCC50071) and B-mode salmonella typhi (preserving number is CMCC50094) are purchased from U.S. ATCC center and China General Microbiological culture presevation administrative center respectively.
This phage STP4-a is stored in China typical culture collection center, address: Wuhan, China, Wuhan University, deposit number: CCTCC NO:M 2014145, taxonomy is named: Salmonellas phage STP4-a (Salmonella bacteriophage STP4-a).
Embodiment 1: phage is separated preparation and purifying
Be seeded to by Salmonella typhimurium ATCC14028 in 10mL nutrient broth, overnight shaking is cultivated, and obtains Host Strains liquid.The sewage sample getting different location, Qingdao of Shandong province next day respectively with Host Strains liquid mixed culture after, centrifugal (3318g, 10min) get supernatant liquor through millipore filtration (0.22 μm) the degerming solution obtained containing phage, get this solution 0.1mL, carry out 10 times of dilutions, the Host Strains liquid 0.1mL getting each 0.1mL of diluent and incubated overnight mixes, add about 5mL semisolid medium, nutrient agar plate upper strata is poured into rapidly after mixing, shake up horizontal 5min, treat that it solidifies, be placed in after 37 DEG C of constant incubators cultivate 12h and observe, obtain the double-layer plate forming plaque, then the purifying that phage is carried out at least 3 single spot punctures is repeated continuously, phage after purifying is joined in Host Strains liquid and carries out multiplication culture, centrifugal (7607g, 20min) supernatant liquor obtains phage stoste after millipore filtration (0.22 μm) is degerming.
Semisolid medium: by 19g/L nutrient broth medium and 33g/L nutrient agar formulated according to the volume ratio of 1:1.
Phage electron microscopic observation
By prophage drop in paraffin plate, copper sheet net is placed in dropping liquid, after 15min, sucks surplus liquid with filter paper, drip phosphotungstic acid aqueous solution, suck surplus liquid after dyeing 10min, dry 10-15min in atmosphere, observes the form of phage under transmission electron microscope.As shown in Figure 1, phage STP4-a is tadpole shape, and head length is 100nm, and transverse diameter is 80nm, has the elongate tail that length is about 120nm, belongs to Myoviridae.
Embodiment 2 temperature and pH are on the impact of phage
EP pipe containing 900 μ L liquid nutrient mediums is put in preheating 60min in the water-bath of differing temps value, after temperature-stable, adds 100 μ L phage stostes, after 60min cultivates, measure phage titer; The liquid nutrient medium of different for 900 μ L pH value is joined in the EP pipe of 2mL respectively, is placed in the constant incubator preheating 30min that temperature is 37 DEG C, after temperature-stable, adds 100 μ L phage stostes, after 60min cultivates, measure phage titer.
As shown in Figure 2, under 40 DEG C of conditions, the activity of phage remains unchanged result in 30min; Under 50 DEG C of conditions, during 15min and 30min, still have the activity of 90% and 75% respectively, but after 60-80 DEG C, 15min process, the activity of phage only remains less than 0.1%.After 30min process, the survival number of phage is lower than 10pfu/mL.
As shown in Figure 3, STP4-a is under pH is the condition of 4-7 to result, and survival rate is 100%.When pH is 8-10, the survival rate of phage still has 90%.When pH is 11 and 12, the survival rate of phage is respectively 70% and 56%.
Embodiment 3 phage preservation condition is analyzed
Utilizing diverse ways preservation phage 1. the phage solution that purifying is good to be added final concentration is under being kept at 4 DEG C and-20 DEG C of conditions after the glycerine of 20%.2. the phage solution that purifying is good being added final concentration is use liquid nitrogen freeze-drying immediately after the skimmed milk of 30%, under being kept at-80 DEG C of conditions.Preserve respectively after 1 year and judge the feasibility of store method by measuring tiring of phage.
As shown in Figure 4 ,-20 DEG C of cryopreservation effects are best for result.4 DEG C and-80 DEG C of cryopreservation phages are after 1 year, and tiring of phage reduces nearly 2 and 1 order of magnitude respectively.
Embodiment 4 phage splitting spectrum analysis
By microbionation to 10mL liquid nutrient medium, shaking culture, to logarithmic phase, is respectively got 100 μ L bacterium liquid and is joined in 5mL semisolid medium, be poured onto in the flat board containing one deck substratum and form double-layer plate, after dry, by 10 of 10 μ L
2to 10
10the phage point that pfu/mL tires drops on flat board, and 37 DEG C of inversions are cultured to plaque and occur.
Result is as shown in table 1, with Host Strains Salmonella typhimurium ATCC14028 for reference, STP4-a can the Salmonellas of all the other 5 strain different serotypes of cracking on an equal basis efficiently, the phage solution of different weaker concn is all defining the high plaque of Clear & Transparent degree containing on the flat board of Salmonellas, and the Salmonellas of display STP4-a to different serotypes has stronger lytic effect.
Table 1 phage splitting spectrum analysis
Remarks :+representative produces transparent circle, namely has cracking ability ,-representative does not produce transparent circle, does not namely have cracking ability.
The Oral toxicity experiment of embodiment 5 Salmonellas phage STP4-a
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 system, the thing tech equipment company limited that supports one's family is praised in Guangzhou.
UNIQ-10 pillar bacterial genomes DNA extraction agent box, Shanghai Sheng Gong biotechnology limited-liability company;
DNase, Beijing Suo Laibao Science and Technology Ltd.;
RNase, Beijing Suo Laibao Science and Technology Ltd.;
Bar-shaped feed, institute for drug control, Qingdao City.
Main agents is prepared
SM damping fluid: accurately take 5.8g NaCl, 2g MgSO
47H
2o is dissolved in 800mL distilled water, adds 50mL 1mol/L Tris-HCl (pH 7.5), adding distil water to 1000mL, 121 DEG C of autoclaving 15min, room temperature preservation.
1mol/L Tris-HCl (pH 7.5) solution: accurately take 121.1g Tris adding distil water to 800mL, regulates pH to 7.5, adds distilled water to 1000mL, 121 DEG C of autoclaving 15min, room temperature preservation.
The Oral toxicity experiment of Salmonellas phage STP4-a
Phage propagation is 1.5 × 10 to tiring
10pfu/mL (solvent is SM damping fluid), is placed in serum bottle and saves backup in 4 DEG C of refrigerators;
Buy BALB/c mouse totally 30 (male mouse 15, female mouse 15), 6-8 week age, body weight 16-22g, is put in Animal House endoadaptation environment one week.After conforming, to every mouse weights before gavage, and mark with 3-5% picric acid.This experiment carries out corresponding disposable gavage process to mouse, and process grouping is in table 2, and before contamination, mouse needs fasting to can't help water 3-4h, needs to weigh to animal after fasting before contamination, still needs to continue fasting 1-2h after contamination.In feeding process, the room temperature of Animal House controls at 22 ± 3 DEG C, and relative humidity is 30-70%.Should adopt source of artificial light (bright+10h of 14h is dark) in receptacle, animal edible standard feed, freely drinks water.
Table 2 different grouping is to the processing mode of mouse
Note: F, M represent female mouse group, male mouse group respectively; 1,2,3 represent respectively blank group, negative control group, phage treatment group.
Should respectively do once careful clinical observation by 30min and 4h after contamination to the same day of animal contaminated, once a day later, Continuous Observation 14d.
(1) body weight: every animal of should weighing after contamination, carries out body weight determination every other day, and carries out body weight record, calculates body weight change;
(2) gross examination of skeletal muscle: observe and should comprise skin, changed by hair, eyes and mucous membrane, also should observe the sign of breathing, circulation, vegetative nerve and central nervous system, limb activity situation and Behavioral change;
(3) histopathologic examination: tackle animal of all contaminating (comprising the animal that duration of test is dead or put to death) and carry out gross anatomy, the weight of record liver, kidney, spleen and thymus gland, calculates organ coefficient (organ coefficient=organ weights/body weight × 100%); Record whole macroscopic disease damage organ (to observe organ and comprise liver, kidney, spleen, stomach, intestines, heart, preserve in 40% formaldehyde solution), tissue sample through draw materials, fix, dewater transparent, waxdip, paraffin embedding, section, phenodin-Yihong (H.E) staining, mounting (Zhao Yongqiang. the generation of free radical and the impact on product quality and security thereof in Tilapia Fillet ozone bacteria reducing process: [Ph.D. Dissertation]. Shandong: Food Science and Engineering institute of Chinese Marine University,, and do histopathologic examination 2013.).
The inspection of data analysing method two group difference adopts t-inspection, and P<0.01 is judged as having pole significant difference, and P<0.05 is judged as having significant difference, and P>0.05 is that difference is not remarkable.Testing data employing mean value ± standard deviation (
) represent.
Experimental result
Blank group, negative control group, each 10 mouse of experimental group are during gavage and in 14 day observation period, without the phenomena of mortality, and respectively organize mouse skin, by hair, eyes and mucous membrane no abnormality seen, breathe, circulation, vegetative nerve and central nervous system sign, limb activity situation and Behavioral change also phenomenon without exception occur.Table 3, table 4, table 5 represent the statistics of each group of Mouse Weight, internal organs weight and organ coefficient respectively.
(1) body weight change
Deactivation phage group, phage group Mouse Weight at each minute point compared with SM buffer control group, through t-check analysis all there was no significant differences (P>0.05), illustrate that phage and residue thereof do not make significant difference to Mouse Weight, the results are shown in Table 3.
Table 3 each group Mouse Weight statistics (g,
)
Note: in table, subscript represents significant difference result, "; " front letter representation F2 and F1 and M2 and M1 comparative result, "; " rear alphabetical F3 and F1 and M3 and M1 comparative result, same letter represents there was no significant difference (P > 0.05), and different letter representation has significant difference (P < 0.05), and "-" expression has neither part nor lot in comparison procedure.
(2) organ weights
Phage group, deactivation phage group are compared with SM buffer control group, through t-check analysis, kidney, spleen, thymic weight there was no significant difference (P > 0.05), illustrate that phage and residue thereof do not make significant difference to mouse kidney, spleen, thymic weight.Wherein, t-check analysis finds, F2 group liver weight is significantly higher than F1 group (P < 0.05), illustrates that the residue of phage can not have a negative impact to liver growth, likely can promote liver development.The results are shown in Table 4.
The heavy result of table 4 each group mice organs (g,
)
Note: in table, subscript represents significant difference result, "; " front letter representation F2 and F1 and M2 and M1 comparative result, "; " rear alphabetical F3 and F1 and M3 and M1 comparative result, same letter represents there was no significant difference (P > 0.05), and different letter representation has significant difference (P < 0.05), and "-" expression has neither part nor lot in comparison procedure.
(3) organ coefficient
Phage group, deactivation phage group are compared with SM buffer control group, through t-check analysis, kidney, spleen, thymus gland coefficient there was no significant difference (P > 0.05), illustrate that phage and residue thereof do not make significant difference to mouse kidney, spleen, thymus gland coefficient.Wherein, t-check analysis finds, F2 group liver weight is significantly higher than F1 group (P < 0.05), illustrates that the residue of phage can not have a negative impact to liver growth, likely can promote liver development.The results are shown in Table 5.
Table 5 each group mice organs coefficient results (%,
)
Note: in table, subscript represents significant difference result, "; " front letter representation F2 and F1 and M2 and M1 comparative result, "; " rear alphabetical F3 and F1 and M3 and M1 comparative result, same letter represents there was no significant difference (P > 0.05), and different letter representation has significant difference (P < 0.05), and "-" expression has neither part nor lot in comparison procedure.
(4) pathological examination result
A. gross anatomy result
All do not find that unusual phenomenon appears in internal organs color, structure etc. after all mouse dissections of blank group, deactivation phage treatment group and phage treatment group.So, often organize random picking mouse making tissue pathological slice and observe whether have unusual phenomenon further.
B. histopathological examination result
No significant difference between the section of F1, F2, F3 group and between the section of M1, M2, M3 group, Fig. 5, Fig. 6, Fig. 7, Fig. 8, Fig. 9, Figure 10 represent the tissue slice figure of different liver, spleen, kidney, stomach, intestines and the heart organized respectively.Wherein liver organization structure is normal, and without extravasated blood, ischemic, oedema, necrosis, fibrosis, tunicle is complete.Liver lobule, central vein is positioned at centrilobular, and tube wall is complete, has liver sinusoid to import.Liver rope, liver cell monolayer alignment is formed, and shows greatly centered by central vein and becomes radial arrangement to surrounding; Spleen tissue structure is normal, without extravasated blood, oedema, hyperplasia; Renal tissue structure is normal, without oedema, necrosis, fibrosis.Visible renal glomerulus in cortex, renal glomerulus structure is normal, and number is normal, visible a large amount of different tangent plane in medullary substance, and intensive uriniferous tubules arranged in parallel, without renal corpuscule; Have no hemorrhage, downright bad in stomach-tissue, rotten to the corn or ulceration; The arrangement of intestinal tissue submucosal glands is still neat, muscle layer and outer membrane structure complete; Heart tissue structure is normal, without downright bad, fibrosis.Cardiac muscle fibre takes on a red color, visible a small amount of loose connective tissue and a large amount of capillary vessel between cardiac muscle fibre.Respectively be organized in the section of M1, M2, M3 group and also can draw same conclusions.From histopathologic slide's result, each group of tissue slice figure is without significant difference, visible phage STP4-a to each organ-tissue without obvious negative effect, 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, the security of phage is all demonstrated from experimentation on animals aspect.
This experiment further illustrates the security of phage STP4-a by Oral toxicity experiment, data analysis find phage treatment group, deactivation phage treatment group with blank group in body weight, organ weights, internal organs growth etc. compared with all there is no significant difference.And by gross anatomy and tissue slice observe do not find mouse take phage after intracorporeal organ there is any pathology phenomenon.From molecular level analysis and Oral toxicity experimental result, phage does not have obvious damaging effect, is a kind of safe antiseptic-germicide.
Embodiment 6: the bacteriostatic experiment of Salmonellas phage STP4-a in laying hen body
36 17 age in days laying hens are divided into 3 groups, as table 6.Except A group, all the other respectively organize every chicken intragastrically oral 10 respectively
7cfu Salmonella typhimurium ATCC14028.Mixed with feed by phage, its concentration is 10
9pfu/g feed, containing phage feed from 3 days feedings before microbiological contamination, feeds 14 days continuously after microbiological contamination again, the 7th day and the 14th day dead 6 chickens in every component other places, takes out its enteron aisle to measure bacterium number.Owing to containing the advantage intestinal microfloras such as intestinal bacteria in chicken intestinal, estimate the quantity considerably beyond Colonization inside plants Salmonellas ATCC14028, so first increase bacterium process through Salmonellas when measuring, after increasing bacterium by mensuration, whether Salmonellas number changing conditions understanding phage STP4-a has remarkable effect effect in chicken intestinal.Measuring method is as follows: by small intestine by 1 (g): 10 (mL) immerse in magnesium chloride Victoria Green WPB (RV) enrichment liquid, after cultivating 24h in 42 DEG C after aseptic homogeneous, sample stroke-physiological saline solution is carried out ten times of gradient dilutions, selecting appropriate dilution to get 100 μ L is applied on xylose lysine deoxidation cholate (XLD) flat board, carries out Salmonellas enumeration after 37 DEG C of cultivation 24h.
Table 6 experimental design
Count the Salmonellas in the chicken intestinal of different treatment group respectively when the 7th day and the 14th day, A group does not all detect Salmonellas, and the Salmonellas thus in B, C group enteron aisle is all oral Salmonellas.In different treatment group, different feeding time chicken intestinal, Salmonellas content results is in table 11, and wherein " * * " shows that phage treatment group and positive controls have pole significant difference, P<0.01; Cubes represents bacterium number in every chicken intestinal, and horizontal line represents the average of bacterium number in this group 6 chicken intestinal.
As shown in Figure 11, the 7th day time, C group Salmonellas number comparatively B group have dropped 0.25log (cfu/g), and through t-check analysis, P>0.05, decline effect is not remarkable.During by 14 days, C group Salmonellas number have dropped 1 order of magnitude compared with B group, and through t-check analysis, extremely significantly (P < 0.01), this illustrates that phage can play certain result for the treatment of in chicken body to decline effect.
Above-described embodiment is one of the present invention preferably scheme, not does any pro forma restriction to the present invention, also has other variant and remodeling under the prerequisite not exceeding the technical scheme described in claim.
Claims (6)
1. a strain wide fragmentation pattern Salmonellas phage, it is characterized in that: preserving number is: CCTCC NO:M 2014145, this phage has effective splitting action to Salmonellas.
2. as described in claim 1, phage, preparing the application killed in the medicine of Salmonellas, is characterized in that: after Salmonellas phage purifying, for suppressing and killing Salmonellas.
3. application according to claim 1, it is characterized in that: this phage is used for preventing salmonella-polluted and Salmonellas hypertrophy, and wherein said Salmonellas comprises Salmonella typhimurium, Salmonella paratyphi A, Salmonella enteritidis, salmonella typhi and B-mode salmonella typhi.
4. the application of wide fragmentation pattern Salmonellas phage according to claim 1 in control food contamination Salmonellas.
5. according to the application described in claim 4, it is characterized in that: after phage dilute with water, as flushing liquor or leacheate, for the production environment of food or produce utensil or production unit carries out sprinkling dissipation, for controlling the pollution of Salmonellas to described environment or utensil or equipment.
6. the application according to claim 3 or 4, is characterized in that: described food refers to: by the fabricated product selecting in grain, meat, vegetables, eggs, or the fabricated product combined by their.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1602353A (en) * | 2001-12-13 | 2005-03-30 | 雀巢产品有限公司 | Isolated phages and their use in food or pet food products |
| CN102041247B (en) * | 2010-10-15 | 2012-11-07 | 江苏省农业科学院 | Salmonella bacteriophage and application thereof |
-
2014
- 2014-09-28 CN CN201410508239.XA patent/CN104830806B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1602353A (en) * | 2001-12-13 | 2005-03-30 | 雀巢产品有限公司 | Isolated phages and their use in food or pet food products |
| CN102041247B (en) * | 2010-10-15 | 2012-11-07 | 江苏省农业科学院 | Salmonella bacteriophage and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 包红朵 等: "宽噬菌谱肠炎沙门氏菌噬菌体的生物学特性", 《江苏农业学报》 * |
| 李萌 等: "一株沙门氏菌烈性噬菌体的分离纯化与生理特性研究", 《水产科学》 * |
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| CN106497888A (en) * | 2016-10-29 | 2017-03-15 | 华中农业大学 | Salmonella phage and phage bactericidal composition and its application |
| WO2019154032A1 (en) * | 2018-02-07 | 2019-08-15 | 青岛诺安百特生物技术有限公司 | Broad spectrum salmonella phage and application thereof |
| CN108546685A (en) * | 2018-04-20 | 2018-09-18 | 华中农业大学 | A kind of Salmonella enteritidis bacteriophage LPSE28 and its application in food |
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| WO2019212244A1 (en) * | 2018-05-03 | 2019-11-07 | 경북대학교 산학협력단 | Bacteriophage-derived recombinant protein having antimicrobial activity against pathogenic gram-negative bacteria |
| 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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| WO2022048061A1 (en) * | 2020-09-05 | 2022-03-10 | 菲吉乐科(南京)生物科技有限公司 | Salmonella phage having high temperature tolerance and wide lysis spectrum and composition thereof |
| CN113150994A (en) * | 2021-04-15 | 2021-07-23 | 上海理工大学 | Low-temperature preservation method for virulent phage |
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