CN113337480B - Broad-spectrum coliphage and application thereof - Google Patents
Broad-spectrum coliphage and application thereof Download PDFInfo
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- CN113337480B CN113337480B CN202110740470.1A CN202110740470A CN113337480B CN 113337480 B CN113337480 B CN 113337480B CN 202110740470 A CN202110740470 A CN 202110740470A CN 113337480 B CN113337480 B CN 113337480B
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Abstract
The invention belongs to the technical field of biology, and particularly relates to a broad-spectrum escherichia coli bacteriophage and application thereof. The coliphage WXLECP008(Escherichia coli phage, WXLECP008) provided by the invention is a virulent phage, the phage has tolerance under the conditions of pH 4-pH 10 and-80-65 ℃, the phage is cultured for 8 hours under the condition that MOI is 0.1, and the titer is 3.5 multiplied by 1010pfu/mL. The coliphage provided by the invention is a broad-spectrum phage with wide cracking spectrum, high in titer and high in safety, can be massively propagated in nonpathogenic host bacteria, and can be used for large-scale propagationAnd (4) large-scale fermentation production. The phage can provide excellent strain resources for developing phage therapy, has good application and development prospects, and is suitable for popularization and application.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a broad-spectrum escherichia coli bacteriophage and application thereof.
Background
Escherichia coli (Escherichia coli) is commonly referred to as Escherichia coli and has been considered a constituent of the normal intestinal flora for a considerable period of time and is considered to be a nonpathogenic bacterium. Until the middle of the 20 th century, it was not recognized that some specific serotypes of E.coli were pathogenic to humans and animals, often causing severe diarrhea and sepsis. Therefore, most E.coli are generally regarded as opportunistic pathogens. Pathogenic e.coli are classified into 6 classes according to different biological properties: enteropathogenic Escherichia coli (EPEC), enterotoxigenic Escherichia coli (ETEC), enteroinvasive Escherichia coli (EIEC), enterohemorrhagic Escherichia coli (EHEC), enteroadhesive Escherichia coli (EAEC), and Diffusible Adhesive Escherichia Coli (DAEC). Coli can be classified into more than 150 types according to the difference of thalli antigens, wherein 16 serotypes of the escherichia coli are pathogenic and often cause epidemic infantile diarrhea and adult costal meningitis. Currently, colibacillosis brings more and more loss to the breeding industry, and clinical characteristics of difficult cure, high mortality rate, easy recurrence and the like are puzzling vast farmers and veterinarians at the basic level. Then how so difficult is colibacillosis eradicated? Antibiotic resistance, secondary or concurrent infection, environmental pollution, and the like are generally considered. Over the past two thirty years, the problem of antibiotic resistance has become very serious, and resistant strains continue to emerge, which poses significant challenges for medical treatment. It was reported that 100000 cases of MARSA (methicillin-resistant Staphylococcus aureus) infection in the United states caused 20000 deaths in 2005. Treatment of mdr (multidug resistant) bacteria, ESKAPE (Enterococcus faecalis, Klebsiella penicilliae, actinobacter baumannii, Pseudomonas aeruginosa, Staphylococcus aureus) pathogenic bacteria not only increases the associated expenditure, but also presents a potential crisis to the health care industry. Therefore, it is imperative to find a new, non-antibiotic substance to treat bacterial infections.
Bacteriophages, also known as bacterial viruses, can infect bacteria, spirochetes, actinomycetes, mycoplasma, cyanobacteria, and the like. It is widely found in soil, air, water and organisms, presumably the most abundant species in biospheres, in almost every corner of the earth. The total number of which is about 1031And the number is almost 10 times of that of the bacteria. The phage can infect bacteria and multiply in the thallus to lyse the bacteria and kill them. As a natural killer of bacteria, the bacteriophage has outstanding superiority in controlling bacterial infection. The positive effect of using phages to treat bacterial infections was achieved as early as the early 20 th century. Since it has a lytic effect on a specific host bacterium, it is considered that the phage can be used as an agent against bacterial infection.
Wangjie (analysis and clinical application of chicken farm coliphage [ D ]. Master academic thesis of Shandong university, 2017) finds that coliphage vB-EcolP-YSF 1 shows a good effect in treating chicken colibacillosis, has a good clinical application value under actual production conditions, and lays a foundation for later-stage phage treatment of avian colibacillosis.
Road construction Biao and the like (separation identification and treatment test [ J ] of a chicken colibacillus phage, Chinese veterinary science of prevention, 2019.) find that the coliphage Bp1805 can crack pathogenic escherichia coli, and provide a reference basis for phage prevention and control of clinical colibacillosis.
Korea song (2017) discloses enterohemorrhagic Escherichia coli bacteriophage and a preparation method of the application thereof, which are used for treating and preventing infectious diseases of chicken Escherichia coli.
In 2006, the FDA in the united states approved a listeria phage that could be used as a food additive to prevent listeria contamination in cheese food; in 2007, the application range of this phage was expanded from cheese food to all food.
In 2007, the FDA granted escherichia coli O157: h7 was marketed as a phage preparation; in the same year, the American Ministry of agriculture approved the application of phage sprayed on the surface of edible cattle for preventing and treating Escherichia coli pollution; clinical phase one trials of phage cocktail therapy comprising coliphage, staphylococcus aureus phage and pseudomonas aeruginosa phage were conducted at Texas wound care center, usa.
These mark the development and application of phages to a new stage. Under such a background, bacteriophage is not only used for treating MDR bacterial infection, but also widely used in various fields such as food, agriculture, livestock husbandry, industry, and clinical diagnosis, vaccine research, etc.
At present, researches on coliphage mainly focus on the aspect of sterilization and bacteriostasis, researches on more pathogenic coliphage and cracking characteristics thereof are still few, how to enrich broad-spectrum phage resources, and finding new phage with strong cracking performance is an urgent problem to be solved in the technical field of prevention and treatment of colibacillus pathogenic bacteria. In conclusion, there is an urgent need to develop novel phages for E.coli with a broad lysis spectrum.
Disclosure of Invention
The invention provides a one-strain broad-spectrum escherichia coli bacteriophage and application thereof, aiming at solving part of problems in the prior art or at least alleviating part of problems in the prior art.
The invention is realized in such a way that a broad-spectrum coliphage with the preservation number of CCTCC M2020658 is preserved in 30 days 10 months 2020 to China. Escherichia coli phage (Escherichia coli phase) WXLECP 008.
The invention also discloses application of the broad-spectrum Escherichia coli bacteriophage in Escherichia coli lysis.
The invention also discloses application of the broad-spectrum escherichia coli bacteriophage in preparation of a medicine for cracking escherichia coli.
Further, the medicament comprises a carrier medicament and/or a liquid medicament.
Further, the carrier medicament comprises an aqueous carrier.
Further, the aqueous carrier comprises a phosphate buffer and/or water.
Further, the liquid medicament disinfectant and/or rinsing agent.
Further, the medicine comprises a preservation number of CCTCC NO: m2020658 phage or its fermentation broth; the preparation method of the fermentation liquor comprises the steps of mixing the phage and the host bacteria, and inoculating the mixture into a TSB culture medium for culture.
The invention also discloses application of the broad-spectrum coliphage in treating sewage containing colibacillus.
The coliphage WXLECP008(Escherichia coli phase, WXLECP008) is a virulent phage which has tolerance at a pH of 4-10 and a temperature of-80-65 ℃, and the titer is 3.5 multiplied by 10, the MOI is 0.1, the phage is cultured for 8h10pfu/mL。
The Escherichia coli phage WXLECP008(Escherichia coli phase, WXLECP008) has the following physiological characteristics and beneficial effects in the aspects of phage titer, optimal infection Multiplicity (MOI) of cracked Escherichia coli, pH stability, temperature tolerance and the like:
(1) has higher titer (see table 1), wherein the optimal MOI of Escherichia coli infected by Escherichia coli phage WXLECP008(Escherichia coli phase, WXLECP008) is 1:10 respectively; is a virulent phage with high affinity and lytic ability, and can rapidly lyse Escherichia coli (Table 2); the growth activity of the coliphage WXLECP008(Escherichia coli phase, WXLECP008) is influenced the least when the pH value is 6-8; the stability at different temperatures is relatively good; is suitable for low-temperature storage at 4 ℃.
(2) Coli phage WXLECP008(Escherichia coli phase, WXLECP008) avirulent or undesirable genes; the tested phage has a wider host range, and the recognition rate of the tested phage on 142 strains of Escherichia coli reaches 82.4%.
(3) Lysis of non-pathogenic bacteria by the E.coli phage WXLECP008(Escherichia coli phase, WXLECP 008): 27 non-pathogenic and beneficial strains including Staphylococcus aureus and enterococcus faecium could not be identified.
(4) The E.coli phage WXLECP008(Escherichia coli phase, WXLECP008) has the following advantages: it is a strictly virulent phage and highly toxic to host bacteria; has a wide host range; can be well proliferated on a non-pathogenic bacterial host; large-scale fermentation culture can be carried out; the culture solution can stably survive at room temperature and can maintain activity for a long time at 4 ℃. The present invention does not subject the test phage to any genetic modification. Therefore, the Escherichia coli phage WXLECP008(Escherichia coli phase, WXLECP008) can provide excellent strain resources for developing phage therapy, and has good application and development prospects.
(5) Animal experiments prove that the coliphage WXLECP008 has a treatment effect on avian colibacillosis and has good capability of bleeding through intestinal tracts.
(6) The coliphage WXLECP008 of the present invention can be prepared into a medicament or a reagent for treating or preventing diseases caused by escherichia coli by those skilled in the art according to the description of the present invention and the common general knowledge in the art.
(7) The coliphage WXLECP008 of the invention includes but is not limited to the range of being applied to the body surface, living water body and the like of a host to be controlled in the forms of carrier carrying, concentrated spraying or medicament soaking and the like; as one embodiment, the carrier-borne form includes, but is not limited to, aqueous carriers; concentrated spray forms include, but are not limited to, sprays and the like; the medicament soaking form includes but is not limited to rinsing agent and the like.
(8) The coliphage WXLECP008 of the invention is prepared into various products which are often used as effective components for environmental disinfection, for example, but not limited to, the products can be used for disinfecting and decontaminating water distribution systems, aquaculture facilities or other environmental surfaces in the forms of liquid soaking, spraying, combined use with aqueous carriers and the like, and the growth and activity of target bacteria can be effectively controlled. The liquid soaking and spraying forms include but are not limited to disinfectants and the like; the aqueous carrier includes, but is not limited to, phosphate buffer, seawater, and the like.
In summary, the advantages and positive effects of the invention are:
the coliphage provided by the invention is a broad-spectrum phage with wide cracking range, has high titer and high safety, can be massively propagated in non-pathogenic host bacteria, and can be used for large-scale fermentation production. The phage can provide excellent strain resources for developing phage therapy, has good application and development prospects, and is suitable for popularization and application.
Drawings
FIG. 1 is a graph of the effect of phages on E.coli lawn;
FIG. 2 is a structural diagram of a phage.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the equipment and reagents used in the examples and test examples are commercially available without specific reference. The specific embodiments described herein are merely illustrative of the invention and are not intended to be limiting.
Various modifications to the precise description of the invention will be readily apparent to those skilled in the art from the information contained herein without departing from the spirit and scope of the appended claims. It is to be understood that the scope of the invention is not limited to the procedures, properties, or components defined, as these embodiments, as well as others described, are intended to be merely illustrative of particular aspects of the invention. Indeed, various modifications of the embodiments of the invention which are obvious to those skilled in the art or related fields are intended to be covered by the scope of the appended claims.
For a better understanding of the invention, and not as a limitation on the scope thereof, all numbers expressing quantities, percentages, and other numerical values used in this application are to be understood as being modified in all instances by the term "about". Accordingly, unless indicated otherwise, the numerical parameters set forth in the specification and attached claims are approximations that may vary depending upon the desired properties sought to be obtained. At the very least, each numerical parameter should at least be construed in light of the number of reported significant digits and by applying ordinary rounding techniques. In the present invention, "about" means within 10%, preferably within 5% of a given value or range.
In the following examples of the present invention, the temperature is not particularly limited, and all of the conditions are normal temperature conditions. The normal temperature refers to the natural room temperature condition in four seasons, no additional cooling or heating treatment is carried out, and the normal temperature is generally controlled to be 10-30 ℃, preferably 15-25 ℃.
The invention discloses a broad-spectrum Escherichia coli bacteriophage and application thereof, wherein the Escherichia coli bacteriophage WXLECP008(Escherichia coli phase, WXLECP008) has a preservation number of CCTCC M2020658, the preservation unit is China center for type culture collection, and the preservation time is 2020.10.30. The formula of the culture medium is as follows:
the formula of the TSB liquid culture medium is as follows: 17g of tryptone, 3g of soybean papain digest, 5g of sodium chloride, 2.5g of monopotassium phosphate, 2.5g of glucose, 1000mL of distilled water and pH 7.0.
The formula of the TSA solid culture medium is as follows: tryptone 15g, soybean papain digest 5g, sodium chloride 5g, agar 15g, distilled water 1000mL, pH 7.0.
The formula of the TSB semisolid agar culture medium is as follows: 17g of tryptone, 3g of soybean papain digest, 5g of sodium chloride, 2.5g of monopotassium phosphate, 2.5g of glucose, 7g of agar, 1000mL of distilled water and pH 7.0.
The SM liquid formula is as follows: 8.5g of sodium chloride, 2g of magnesium sulfate, 50mL of 1mol/L Tris-HCl, 0.25g of gelatin and 1000mL of distilled water.
Example 1 E.coli phage WXLECP008 isolation and purification
Collecting a certain place in Hubei10 portions of sewage samples near the chicken farm, each portion is 20mL, 10mL of supernatant is taken after centrifugation for 10min at 5000rpm, the supernatant is sterilized, and the supernatant is mixed with 10mL of 2 xTSB liquid culture medium and 2mL of escherichia coli WXLEC008 bacterial liquid (10 mL) in logarithmic phase8cfu/mL) were mixed uniformly and cultured overnight at 37 ℃ at 180rpm to enrich phage. Centrifuging the sample enrichment solution at 5000rpm for 10min, filtering the supernatant with 0.22 μm microporous membrane, and sterilizing to obtain filtrate containing bacteriophage. Taking 100 mu L of filtrate, uniformly mixing with 300 mu L of host escherichia coli liquid, and standing for 15min to ensure that the filtrate is fully combined with receptors on the surfaces of bacteria. And adding the mixed solution into 4mL of TSB semisolid agar culture medium cooled to 50 ℃, uniformly mixing, immediately paving on a solidified TSA plate, after the agar is solidified, carrying out inverted culture at 37 ℃ for 8-12h, and observing the growth condition of the plaque. Picking large and transparent plaques on a double-layer plate with the plaques, shaking and desorbing the plaques in 1mL of SM solution by using an aseptic gun head, sterilizing the plaques by using a 0.22-micron microporous filter membrane to obtain phage filtrate, inoculating the phage filtrate into 5mL of TSB liquid culture medium, adding 0.1mL of escherichia coli liquid, uniformly mixing, shaking and culturing at 37 ℃ overnight at 180rpm, centrifuging at 5000rpm for 10min, taking supernatant, filtering by using a bacterial filter membrane, and observing the plaque shape by using a double-layer plate method. Repeating the operation for 3-5 times to obtain plaques with consistent shape and size. And E.coli phage WXLECP008 is separated from the Hubei chicken farm sewage sample. The coliphage WXLECP008 generates single round plaques on the colibacillus lawn, the center is clear, the edge is provided with a halo, and the diameter is 1-2mm (as shown in figure 1); specifically, Myoviridae, the head is about 94X 44nm and the tail is about 100X 19nm (FIG. 2).
Example 2 determination of the E.coli phage WXLECP008 titre
Stock E.coli phage WXLECP008 (from example 1) was diluted 10-fold stepwise to l0 using SM as diluent8And (4) doubling. Respectively taking l05、l06、l07And l080.1mL of diluted phage culture solution is uniformly mixed with 500 mu L of host bacteria (avian nonpathogenic Escherichia coli (ANPEC)) Escherichia coli WXLEC008 bacterial solution, and the mixture is kept stand for 15min to be fully combined with receptors on the surfaces of the bacteria. The mixture was added to 4mL of TSB half cooled to 50 deg.CMixing with solid agar culture medium, spreading on solidified solid agar plate, and culturing at 37 deg.C for 8-12 hr. Three replicates of each dilution were taken and counted as the average of the three replicates of the dilution. Wherein, the titer of the phage (PFU/mL) is the average plaque number multiplied by the dilution multiple
As can be seen from Table 1, the isolated E.coli phage WXLECP008 had a 10-degree sequence8Titers above PFU/mL.
TABLE 1 titer of coliphage WXLECP008
| Bacteriophage | WXLECP008 |
| Potency (PFU/mL) | 5.6x108 |
Example 3 determination of the optimal multiplicity of infection (MOI) of E.coli by the E.coli phage WXLECP008
And (3) selecting a single escherichia coli colony, inoculating the single escherichia coli colony into a bacteria bottle containing 3mL of TSB culture solution, and performing shaking culture in a shaker at 37 ℃ for 12h at 180rpm to obtain a host bacteria suspension. The bacterial suspension is transferred to l0mL TSB culture solution in a ratio of 1:100, and is subjected to shaking culture at 37 ℃ and 180rpm until the prophase of logarithm. Pure culture broth of phage WXLECP008 (prepared from example 1) and host bacteria (MOI ═ phage number/bacterial number) were added in terms of the multiplicity of infection ratio, and TSB broth was added to make the total volume of each tube the same. Shaking overnight at 180rpm in a shaker at 37 ℃. After the culture, centrifugation is carried out for l0min at 5000g, and the supernatant is collected to determine the titer of the phage. Each point was subjected to duplicate multi-tube culture and averaged to obtain the MOI producing the highest phage titer as the optimal multiplicity of infection. The experiment was repeated 3 times.
The results are shown in Table 2, the E.coli phage WXLECP008 titer reached the highest (5.3X 10)9PFU/mL), the MOI is 1: 10.
TABLE 2 titer of E.coli phage WXLECP008 at different multiplicity of infection
Example 4 pH stability test of E.coli phage WXLECP008
Adding sterile EP tube into TSB culture medium of different pH (3-11) 900uL, placing the EP tube in thermostatic water bath of 25 deg.C, adding 00 μ L thallus pure culture solution after temperature balancing, and standing at room temperature for 1 h. After the reaction time is over, the sample is diluted properly and then the titer of the phage is measured by adopting a double-layer plate method. The above process was repeated after 1h, 2h, 4h and 24h, respectively, and the experiment was repeated 3 times.
As a result, as shown in Table 3, the E.coli phage WXLECP008 showed no significant change in titer after 24 hours treatment at pH 5-10.
TABLE 3 stability of the values of E.coli phage WXLECP008pH after different times of reaction (initial titer: 5.0X 10)9PFU/mL)
Note: ND means not detected.
Example 5 temperature sensitivity test of E.coli phage WXLECP008
Each 00 μ L phage pure culture solution was filled in sterile EP tube, and applied in water bath at 4 deg.C, 45 deg.C, 55 deg.C, 65 deg.C, 75 deg.C, 85 deg.C, 80 deg.C and-20 deg.C for 1h, 2h, 4h and 12 h. After the action time is over, the sample tube is taken out, the phage titer is measured by adopting a double-layer plate method after the sample tube is properly diluted, and the experiment is repeated for 3 times.
The results are shown in table 4, the coliphage WXLECP008 in the experimental group can survive at 65 ℃, which indicates that the coliphage WXLECP008 has good thermal stability and still has higher titer after being subjected to water bath at 65 ℃ for 1 hour; the high titer is still obtained after the treatment for 12h at the extreme temperature of minus 80 ℃ and minus 20 ℃, which shows that the coliphage WXLECP008 has good low-temperature resistance.
TABLE 4 titer of E.coli phage WXLECP008 at different temperatures (initial titer: 3.5X 10)9PFU/mL)
Example 6 E.coli phage WXLECP008 virulence gene or undesirable gene deletion detection assay
65 identified virulence genes from lysogenic phages derived from pathogenic bacteria (Table 5) were selected and determined to contain the virulence genes by determining the whole genome of E.coli phage WXLECP008 and performing bioinformatics analysis on it. The results show that the test phage did not contain the following virulence genes. The test phage had no undesirable genes.
TABLE 5 major known virulence genes of lysogenic phages in pathogenic bacteria
Example 7 E.coli phage WXLECP008 on the E.coli lytic Range test
Taking the titer of 1x108PFU/mL E.coli phage WXLECP008 stock solution, the lysis profile of the phage was determined by the titration method.
Single colonies of 142 escherichia coli strains are picked and inoculated into test tubes containing 3mL of TSB respectively, and cultured for 8h at 180rpm to prepare bacterial liquid of each strain. mu.L of the bacterial suspension was mixed with the semi-solid medium and plated on a TSA plate, and 10. mu.L of the phage culture solution was dropped on the plate. After natural air drying, culturing for 8-12h at 37 ℃, and observing the result. The experiment was repeated three times.
The results are shown in Table 6, the E.coli phage WXLECP008 has a broad host range. WXLECP008 can crack 117 of 142 Escherichia coli strains, and the cracking rate is 82.4%.
TABLE 6 results of lysis Spectroscopy determination of E.coli phage WXLECP008
Note: "-" indicates no cleavage; "+" indicates slight lysis and blurred lysis spots; "+ +" indicates cleavage, the cleavage spots are more distinct; "+ + + +" indicates cleavage, the cleavage spots are very clear.
Example 8 lysis test of the E.coli phage WXLECP008 against non-pathogenic bacteria
And (3) selecting 27 strains of nonpathogenic bacteria including nonpathogenic escherichia coli, enterococcus faecium, staphylococcus epidermidis and the like, respectively inoculating the single colonies into test tubes containing 3mL of TSB, and culturing at 180rpm for 8 hours to obtain bacterial liquid of each strain. 300. mu.L of each bacterial suspension was mixed with the semi-solid medium and plated on a common agar plate. Each 8. mu.L of phage culture solution was dropped onto the plate. After natural air drying, culturing for 8-12h at 37 ℃, and observing the result. The experiment was repeated three times.
As shown in Table 7, in this study, the Escherichia coli phage WXLECP008 cleaved only 2 strains of Escherichia coli among 27 non-pathogenic bacteria such as 27 non-pathogenic bacteria.
TABLE 7 lysis test of the E.coli phage WXLECP008 against non-pathogenic bacteria
Note: "-" indicates no cleavage; "+" indicates slight lysis and blurred lysis spots; "+ +" indicates cleavage, the cleavage spots are more distinct; "+ + + +" indicates cleavage, the cleavage spots are very clear.
Example 9 fermentative preparation of Escherichia coli phage WXLECP008
Selecting a single colony of escherichia coli WXLEC008, inoculating the single colony into a test tube containing 3mLTSB (in a culture solution, in a shaking table at 37 ℃, carrying out shaking culture at 180rpm for 12h to obtain a host bacterial suspension, transferring the bacterial suspension into 500mL of TSB culture solution according to a ratio of 1:10, carrying out shaking culture at 37 ℃ and 180rpm to a logarithmic prophase, measuring the concentration of the bacterial suspension, inoculating the escherichia coli phage WXLECP008 into a system of 5L in fermentation preparation, using a fermentation medium as the TSB culture medium, adopting a flame inoculation method for inoculation, and respectively inoculating 50mL of phage (10 mL) into the fermentation culture medium according to the corresponding optimal infection complex number ratio8PFU/mL) and host bacterial liquid (10)9CFU/mL). And introducing sterile air in the fermentation process, and adding 3% of defoaming agent, wherein the fermentation preparation time is 12 h. 20mL of a mixture of the phage and the host bacteria was taken from the sampling port every 2 hours from the start of the fermentation, centrifuged at 5000rpm for 10min, and the supernatant was sterilized by a 0.22 μm microfiltration membrane to obtain a filtrate containing the phage and the titer of the filtrate was measured by the method described in example 2. And after the fermentation is finished, taking out all mixed liquor of the phage and the host bacteria from a sampling port, inoculating into a sterile container, centrifuging at 5000rpm for 10min, taking supernatant, carrying out suction filtration by using a vacuum air pump into a sterile filter device to obtain phage fermentation liquor, and storing at 4 ℃.
As can be seen from Table 8, the titer reached the highest when E.coli phage WXLECP008 was fermented for 8 hours,is 3.5x1010PFU/mL. The phage titer is 10 from the initial value after 12h fermentation6PFU/mL increased to 1010PFU/mL, improved by 4 orders of magnitude. Therefore, it is feasible to industrially produce phages on a large scale by fermentation.
TABLE 8 fermentation kinetics of E.coli phage WXLECP008
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (8)
1. A broad-spectrum coliphage with preservation number of CCTCC M2020658.
2. Use of a broad spectrum of coliphage according to claim 1 in the preparation of a medicament for lysing e.
3. Use according to claim 2, characterized in that: the medicament comprises a carrier medicament and/or a liquid medicament.
4. Use according to claim 3, characterized in that: the carrier drug comprises an aqueous carrier.
5. Use according to claim 4, characterized in that: the aqueous carrier comprises a phosphate buffer and/or water.
6. Use according to claim 3, characterized in that: the liquid medicament includes a disinfectant and/or a rinsing agent.
7. Use according to claim 2, characterized in that: the medicine comprises a preservation number of CCTCC NO: m2020658 phage or its fermentation broth; the preparation method of the fermentation liquor comprises the steps of mixing the phage and the host bacteria, and inoculating the mixture into a TSB culture medium for culture.
8. The use of a broad spectrum of coliphage of claim 1 in the treatment of wastewater containing coliform bacteria.
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