A Klebsiella pneumoniae phage capable of lysing multiple K-type K-type Klebsiella pneumoniae, exhibiting long-lasting antibacterial activity, and its applications.

Abstract

The present invention provides a Klebsiella pneumoniae phage that can lyse a variety of K-capsule Klebsiella pneumoniae and has a long-lasting antibacterial effect, and its application, which belongs to the field of biotechnology. The phage was deposited in the China Center for Type Culture Collection on March 4, 2024, and the deposit address is: Wuhan University, Wuhan, China, and the deposit number is CCTCC NO: M 2024373. The phage GRNKpP10 has a lytic effect on Klebsiella pneumoniae. In addition, the phage GRNKpP10 has lytic properties on a variety of K-capsule Klebsiella pneumoniae, and has a long-lasting antibacterial effect on a variety of K-type antigenic Klebsiella pneumoniae; the phage GRNKpP10 has the advantages of high value-added titer, safety and efficiency, and good tolerance to temperature and pH. The present invention can effectively prevent diseases caused by pathogenic Klebsiella pneumoniae in various environments.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, in particular to a Klebsiella pneumoniae phage capable of lysing multiple K-capsule Klebsiella pneumoniae and having long-lasting bacteriostasis, and an application thereof. Background Art

[0002] Klebsiella pneumoniae (Kp) is the most important Gram-negative bacterium in the genus Klebsiella of the family Enterobacteriaceae (commonly known as pneumonia bacillus), and the diseases caused by it account for more than 95% of Klebsiella infections. Klebsiella pneumoniae is a zoonotic pathogen that can cause a series of infections in animals and humans, including respiratory tract, urinary tract, soft tissue and blood infections. It is a new global disease and the cause of purulent liver abscess complicated by metastatic meningitis and endophthalmitis.

[0003] Klebsiella pneumoniae has a variety of virulence factors, including siderophores, pili, capsular polysaccharides (CPS), lipopolysaccharide (LPS), etc. CPS is considered to be the most important virulence factor in Kp. According to the typing of capsular polysaccharides (K antigen), Klebsiella pneumoniae can be divided into 82 K serotypes. Among them, K1, K2 and K5 serotypes are closely related to serious infectious diseases in humans and animals. K1 strains usually cause Friedlander's pneumonia, especially the complication of suppurative liver abscess. K2 and K5 serotypes usually cause community-acquired pneumonia. Rhinosclerosis is associated with K3 serotype. Most of the Klebsiella pneumoniae subspecies isolated clinically from atrophic rhinitis come from K4 serotype and very few K5 serotypes. K54 and K57 strains are associated with the onset of group invasive liver abscess syndrome. Compared with other serotypes, K1, K2, K5, K20, K54 and K57 are highly virulent strains and are commonly found in pneumonia cases. In particular, K1 and K2 are the most virulent, and their prevalence has become increasingly severe in recent years. They have become the main pathogens of clinical bacterial liver abscess syndrome, a phenomenon first discovered in Asia and has now been confirmed around the world. The strong virulence of K1, K2, K54 and K57 may be due to the stronger toxicity of their capsular polysaccharides in the body. Bacterial resistance has always been an important issue in the field of global public health. In addition, the rapid spread of carbapenems and the multidrug resistance of Klebsiella pneumoniae have also attracted much attention.

[0004] At present, the prevention and control of Klebsiella pneumoniae is mainly treated with antibiotics, but the drug resistance of Klebsiella pneumoniae is becoming more and more serious, and the efficacy has certain limitations. Therefore, it is necessary to develop a green, safe, targeted, and long-lasting bacteriophage product for Klebsiella pneumoniae; based on this, the present invention provides a Klebsiella pneumoniae phage that can lyse multiple K-capsule Klebsiella pneumoniae and has long-lasting bacteriophage inhibition and its application. Summary of the invention

[0005] In order to prevent diseases or breeding problems caused by Klebsiella pneumoniae and reduce the morbidity caused by the problems, one of the objects of the present invention is to provide a bacteriophage that inhibits Klebsiella pneumoniae.

[0006] The second object of the present invention is to provide the use of Klebsiella pneumoniae phage GRNKpP10 in the preparation of a drug for treating or preventing diseases caused by Klebsiella pneumoniae infection.

[0007] In order to achieve the above-mentioned purpose, the present invention is realized through the following technical scheme: a Klebsiella pneumoniae phage that can lyse multiple K-capsule Klebsiella pneumoniae and has long-lasting antibacterial effect. The classification and name of the phage is Klebsiella pneumoniae phage GRNKpP10. The applicant isolated a Klebsiella pneumoniae phage GRNKpP10 from the water body in the surrounding sea area of ​​a certain place in Guangdong Province. The phage was sent to the China Center for Type Culture Collection for preservation on March 4, 2024. The preservation address is: Wuhan University, Wuhan, China; the preservation number is CCTCC NO: M 2024373.

[0008] Furthermore, it also includes a complex of Klebsiella pneumoniae phage GRNKpP10, and the phage complex includes but is not limited to a complex of Klebsiella pneumoniae phage GRNKpP10 with other phages and other probiotics.

[0009] An application of the Klebsiella pneumoniae phage GRNKpP10 or phage complex as described above is applied in industries such as environmental disinfection, animal husbandry, poultry farming, aquaculture and biomedicine to prepare an antibacterial agent for Klebsiella pneumoniae.

[0010] Furthermore, the invention is also used for preparing medicines for treating or preventing diseases caused by Klebsiella pneumoniae infection.

[0011] Furthermore, the invention is also used for preparing feed or feed additives for antagonizing Klebsiella pneumoniae.

[0012] Furthermore, the Klebsiella pneumoniae is K1 capsule type, K11 capsule type, K22 capsule type, K24 capsule type, K31 capsule type, K37 capsule type and K64 capsule type Klebsiella pneumoniae.

[0013] Beneficial effects of the present invention:

[0014] 1. The present invention screened out for the first time a Klebsiella pneumoniae phage GRNKpP10 that can lyse multiple K-capsule Klebsiella pneumoniae and has a long-lasting antibacterial effect. The phage GRNKpP10 has a high lysis rate for Klebsiella pneumoniae. The phage GRNKpP10 is effective in inhibiting Klebsiella pneumoniae of different capsule types, and has great reference significance for the prevention, control and treatment of Klebsiella pneumoniae in the future.

[0015] 2. The Klebsiella pneumoniae phage GRNKpP10 provided by the present invention has a high fermentation rate. When the temperature is 28°C and the optimal infection multiplicity MOI is 0.001, the titer can reach 8.8×10 10 PFU / mL; in the industrial fermentation process, the fermentation titer can reach 7.5×10 10 PFU / mL.

[0016] 3. The Klebsiella pneumoniae phage GRNKpP10 provided by the present invention has good environmental tolerance, and the potency of the Klebsiella pneumoniae phage GRNKpP10 does not change significantly in the order of magnitude after being treated in the pH range of 4-11 for 24 hours.

[0017] 4. The Klebsiella pneumoniae phage GRNKpP10 provided by the present invention has no obvious order of magnitude difference in the titer after 16 weeks at 4°C, no obvious order of magnitude difference in the titer after 12 weeks at 25°C, and no obvious order of magnitude difference in the titer after 2 weeks at 37°C; it has good stability at 4-37°C and can be effectively preserved.

[0018] 5. The Klebsiella pneumoniae phage GRNKpP10 provided by the present invention can lyse various K-capsule Klebsiella pneumoniae (K1 capsule type, K11 capsule type, K22 capsule type, K24 capsule type, K31 capsule type, K37 capsule type and K64 capsule type Klebsiella pneumoniae); the Klebsiella pneumoniae phage GRNKpP10 has persistent antibacterial properties and can effectively inhibit the growth of Klebsiella pneumoniae for 24 hours.

[0019] 6. The Klebsiella pneumoniae phage GRNKpP10 provided by the present invention can effectively treat the problem of ascites of largemouth bass caused by Klebsiella pneumoniae. The survival rate of largemouth bass was effectively improved in both the prevention and treatment groups. The survival rate of the blank group was 100%, and the survival rate of largemouth bass in the challenge group was 20%; the survival rate of the treatment group was 85%, the survival rate of the prevention group was 90%, and the survival rate of the enrofloxacin group was 85%. It provides important reference value for the prevention and treatment of Klebsiella pneumoniae in the future, and can be used as a biological antibacterial agent for the prevention and treatment of Klebsiella pneumoniae diseases in largemouth bass and other animals. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 The plaque is of Klebsiella pneumoniae phage GRNKpP10;

[0021] Figure 2 is the titer of Klebsiella pneumoniae phage GRNKpP10 under different infection multiplicity conditions;

[0022] Figure 3 This is a test diagram of the stability of Klebsiella pneumoniae phage GRNKpP10 at different pH values;

[0023] Figure 4 This is a test diagram of the stability of Klebsiella pneumoniae phage GRNKpP10 at different temperatures;

[0024] Figure 5 Schematic diagram of the fermentation dynamics of Klebsiella pneumoniae phage GRNKpP10;

[0025] Figure 6 Schematic diagram of the antibacterial persistence of Klebsiella pneumoniae phage GRNKpP10. DETAILED DESCRIPTION

[0026] In order to make the technical means, creative features, objectives and effects achieved by the present invention easy to understand, the present invention is further explained below in conjunction with specific implementation methods.

[0027] See also Figures 1 to 6 , the present invention provides the following technical solutions:

[0028] Embodiment 1:

[0029] Isolation and Purification of Klebsiella pneumoniae Phage GRNKpP10

[0030] Thirty samples of sewage were collected from a certain place in Guangdong Province, each with 20 mL. The sewage was centrifuged at 6000 rpm for 10 min, and the supernatant was sterilized with a 0.22 μm filter. 10 mL of filtrate, 10 mL of double TSB medium and 1 mL of Klebsiella pneumoniae (concentration of about 5×10 8 CFU / mL) were mixed evenly and cultured overnight in a shaker at 37°C at 200 rpm. The fermentation broth was centrifuged at 6000 rpm for 10 min, the supernatant was sterilized with a 0.22 μm filter, and 100 μL of the filtrate was mixed with 300 μL of Klebsiella pneumoniae (concentration of about 1×10 8CFU / mL) and mix well, add TSB semi-solid (containing 0.65% agar), gently invert and pour evenly on the TSA bottom plate, and after solidification, place the culture dish in a 37°C incubator for overnight culture. Pick up the transparent and bright plaque spots, shake and desorb in 1mL SM liquid, and pass through a 0.22μm microporous filter membrane for sterilization to obtain the phage filtrate, inoculate it in 5mL TSB liquid culture medium, add 500μL of the corresponding host Klebsiella pneumoniae liquid and mix well, shake and culture at 200rpm overnight at 37°C, centrifuge at 6000rpm for 10min, take the supernatant and pass through a 0.22μm microporous filter membrane for sterilization to obtain the phage filtrate, and observe the plaque morphology by double-layer plate method. After repeating the operation 3-5 times, plaques of uniform shape and size can be obtained.

[0031] A Klebsiella pneumoniae phage GRNKpP10 was isolated from the water sample. Figure 1 As shown, the plaques formed by it and the host bacteria are transparent, round, 1.0 mm in diameter, and have obvious lysis zones. The phage was sent to the China Center for Type Culture Collection on March 4, 2024, and the classification name is: Klebsiella pneumoniaephage GRNKpP10, address: Wuhan University, Wuhan, China, and the collection number is CCTCC NO: M 2024373.

[0032] Embodiment 2:

[0033] Determination of the titer of Klebsiella pneumoniae phage GRNKpP10 under different infection multiplicity conditions

[0034] The host bacteria of Klebsiella pneumoniae phage GRNKpP10, Klebsiella pneumoniae KP048, was inoculated into an EP tube with 3 mL TSB and cultured at 37°C for 5-8 hours to obtain KP048 bacterial solution. The bacterial solution was inoculated into 100 mL TSB medium at a 1.5% inoculum, and when KP048 grew to the initial logarithmic phase, phage GRNKpP10 was inoculated. The infection was carried out according to different multiplicity of infection (MOI; MOI = number of phages / number of bacteria, see Figure 2 Each experiment was performed in triplicate. The temperature was set at 28°C and 35°C, and the cells were shaken at 200 rpm for 12 h.

[0035] Results Figure 2 When the temperature was 28°C and MOI = 0.001, the fermentation titer reached a maximum of 8.8 × 10 10 PFU / mL.

[0036] Embodiment 3:

[0037] pH stability test of Klebsiella pneumoniae phage GRNKpP10

[0038] 0.1 mL of Klebsiella pneumoniae phage GRNKpP10 fermentation broth was added to 9.9 mL of TSB liquid culture medium with pH values ​​of 2.0, 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0, 10.0, 11.0, and 12.0, respectively (the titer after dilution was 5*10 7 pfu / mL), put it in a 25℃ incubator for 24h, then take a sample, adjust the pH to 7.4 of the original culture medium with hydrochloric acid and sodium hydroxide solution respectively, make multiple dilutions, and determine the phage titer with the double-layer plate method; duplicate culture was performed at each point to get the average value, and the experiment was repeated 3 times.

[0039] The results are as follows Figure 3 As shown, the titer of Klebsiella pneumoniae phage GRNKpP10 did not change significantly in the order of magnitude when treated with pH in the range of 4-11 for 24 hours.

[0040] Embodiment 4:

[0041] Temperature stability test of Klebsiella pneumoniae phage GRNKpP10

[0042] The Klebsiella pneumoniae phage GRNKpP10 fermentation broth was centrifuged and filtered, and 1 mL was dispensed into 1.5 mL centrifuge tubes (initial titer was 5*10 7 pfu / mL). The centrifuge tubes containing phages were placed in a constant temperature of 4, 25, 37, 45, 55, 65, and 75°C for 0h, 1h, 24h, 48h, 92h, 1w, 2w, 4w, 8w, 12w, and 16w. After the action time, the sample tubes were taken out and immediately placed in an ice bath for cooling. After appropriate dilution, the phage titer was determined by the double-layer plate method. Each point was cultured in duplicate to obtain the average value, and the experiment was repeated 3 times.

[0043] The results are as follows Figure 4 As shown, the survival time of Klebsiella pneumoniae phage GRNKpP10 is longer at temperatures of 45°C and below. At 4°C, the phage titer has no obvious order of magnitude difference after 16 weeks, at 25°C, the phage titer has no obvious order of magnitude difference after 12 weeks, and at 37°C, the phage titer has no obvious order of magnitude difference after 2 weeks. It has good stability at 4-37°C and can be effectively preserved.

[0044] Embodiment 5:

[0045] Host range of bacteriophage GRNKpP10

[0046] The Klebsiella pneumoniae selected in the experiment is divided into 12 K-type capsules, namely K1, K2, K3, K5, K11, K22, K24, K26, K31, K37, K38 and K64. Five strains of each type of Klebsiella pneumoniae were inoculated into 3mL TSB EP tubes, and the tubes were shaken at 37℃ and 200rpm for 8h to obtain Klebsiella pneumoniae liquid. A sufficient amount of TSA bottom plate was prepared, 0.5ml Klebsiella pneumoniae liquid was mixed with 6mL TSB semi-solid medium (containing 0.65% agar) at about 50℃, and then slowly spread on the TSA bottom plate. After the plate medium solidified to room temperature, 5×10 8 5 μL of phage GRNKpP10 with a concentration of PFU / ml was dropped onto the plate, and after air drying, the plate was inverted and placed in a 37°C constant temperature biochemical incubator for about 8 hours, and the results were observed. Each group of experiments was repeated 3 times.

[0047] As shown in Table 1, the Klebsiella pneumoniae phage GRNKpP10 can lyse all K1, K11, K22, K24, K31, K37 and K64 capsule types of Klebsiella pneumoniae; but cannot lyse K2, K3, K5, K26 and K36 capsule types of Klebsiella pneumoniae.

[0048] The experimental results confirmed that the phage GRNKpP10 has a very outstanding effect in dealing with the lytic properties of one or more of the K1 capsule type, K11 capsule type, K22 capsule type, K24 capsule type, K31 capsule type, K37 capsule type and K64 capsule type Klebsiella pneumoniae, which is of great value for the targeted prevention, control and treatment of Klebsiella pneumoniae in the future.

[0049] Table 1 Lysis effect of bacteriophage GRNKpP10 on different capsular types of Klebsiella pneumoniae

[0050]

[0051]

[0052] "-" means no lysis, "+" means lysis and the plaques are brighter; "++" means lysis and the plaques are brighter.

[0053] Embodiment 6:

[0054] Fermentation preparation of Klebsiella pneumoniae phage GRNKpP10

[0055] A single colony of Klebsiella pneumoniae KP048 was picked from the plate, inoculated into 3 mL of TSB medium, and cultured at 37°C and 200 rpm for 8 h to obtain a host bacterial solution. The bacterial suspension was inoculated into 120 mL of TSB medium at a 2% inoculum, and cultured at 37°C and 150 rpm until the early logarithmic phase, when the bacterial solution OD600 value was 0.1.

[0056] Industrial fermentation is a 10L fermenter, a fermentation system is 6L, the culture medium used is TSB, the fermentation temperature is 28°C, the initial pH value is 7.4, a flame method is used to inoculate Klebsiella pneumoniae at 1% inoculum, phage GRNKpP10 is added to the fermentation system at the optimal infection condition MOI=0.001, sterile air is introduced during the fermentation process, and 3‰ defoamer is added, and the fermentation preparation time is 12h. From the beginning of fermentation, 20ml of fermentation liquid is taken from the sampling port in a sterile container at 1h, 2h, 3h, 4h, 5h, 6h, 7h, 8h, 9h, 10h, 11h, and 12h, and centrifuged at 6000rpm for 10min, and the supernatant is sterilized through a 0.22μm microporous filter membrane to obtain a filtrate containing phage and measure its titer, and the method is referenced to Example 2.

[0057] After the fermentation is completed, the entire mixed solution of phage and host bacteria is taken out from the sampling port and placed in a sterile container, centrifuged at 6000 rpm for 10 min, and the supernatant is filtered through a vacuum pump into a sterile filter device to obtain the phage fermentation solution and stored at 4°C.

[0058] Depend on Figure 6 It can be seen that the titer of Klebsiella pneumoniae phage GRNKpP10 reached a maximum of 7.5×10 10 PFU / mL, in the period of 6h to 12h, the titer is stable at 10 10 PFU / mL; The results show that it is feasible to use fermentation method for industrial production of bacteriophages, which provides effective reference data for the preparation of bacteriophage products.

[0059] Embodiment 7:

[0060] Experimental study on the prevention and treatment of Klebsiella pneumoniae phage GRNKpP10 against Klebsiella pneumoniae infection in largemouth bass ascites

[0061] Largemouth bass (Micropterus salmoides), also known as California bass, is an economically valuable freshwater fish. Klebsiella pneumoniae is a conditionally pathogenic bacterium of largemouth bass, which can cause ascites. The diseased largemouth bass is slow to react, and the autopsy symptoms are obvious enteritis and ascites.

[0062] The experimental largemouth bass was provided by a largemouth bass farm in Jingzhou, Hubei. The selected largemouth bass had a normal and healthy appearance, with a size of about 40 / 500g. Before the experiment, the largemouth bass were uniformly placed in a large aquarium to adapt to breeding for 7 days, the water pH was 7.3±0.1, the temperature was 25°C, and the dissolved oxygen was 6.0±0.1mg / L. When the largemouth bass mortality rate was stabilized within 0% to 5% within a week, the experiment was prepared. The Klebsiella pneumoniae used in this embodiment is KP002, and the selected bacteriophage is the Klebsiella pneumoniae bacteriophage GRNKpP10 of the present invention.

[0063] The experimental design was as follows: Largemouth bass were randomly divided into 5 groups, with 20 largemouth bass in each group. The experiment lasted for 7 days:

[0064] 1) Control group: Each largemouth bass was injected with an equal volume of saline (abdominal injection);

[0065] 2) Challenge group: Each largemouth bass was injected with 5×10 9 CFU / mL bacterial solution 100μL (final concentration is 5*10 8 CFU / tail).

[0066] 3) Treatment group: Each largemouth bass was injected with 5×10 9 CFU / mL of bacterial solution 100μL; after 2h, inject 1×10 phage GRNKpP10 7 PFU / treatment only;

[0067] 4) Prevention group: 1 day before the experiment, 1×10 7 PFU / each largemouth bass was injected with 5×10 9 CFU / mL bacterial solution 100μL;

[0068] 5) Enrofloxacin treatment group: Each largemouth bass was injected with a final concentration of 5×10 9 CFU / mL of bacterial solution 100 μL. After 24 hours, add enrofloxacin according to the dosage specified for treatment.

[0069] The experiment lasted for 7 days, and the symptoms and number of deaths of largemouth bass were recorded. The survival rate of largemouth bass in each group was calculated as shown below.

[0070]

[0071] ATotal: The total number of largemouth bass in each group of the experiment, pieces;

[0072] A Death: The total number of largemouth bass deaths in each group of the experiment, pieces;

[0073] C: Survival rate of largemouth bass in each group of the experiment, %.

[0074] The experimental results are shown in Table 2. The survival rate of the blank group was 100%, and the survival rate of the largemouth bass in the challenge group was 0%; the survival rate of the treatment group was 85%, the survival rate of the prevention group was 90%, and the survival rate of the enrofloxacin group was 90%. The experimental results revealed that the Klebsiella pneumoniae phage GRNKpP10 can effectively treat the ascites of largemouth bass caused by Klebsiella pneumoniae. Both the prevention and treatment groups effectively improved the survival rate of largemouth bass. This example provides an important reference value for the prevention and treatment of Klebsiella pneumoniae in the future, and can be used as a biological antibacterial agent for the prevention and treatment of Klebsiella pneumoniae diseases in largemouth bass and other animals.

[0075] Table 2 The control effect of bacteriophage GRNKpP10 on Klebsiella pneumoniae in largemouth bass

[0076] Experimental Group Blank Group Anti-virus group Treatment Group Prevention Group Enrofloxacin group Survival rate 100％ 20％ 85％ 90％ 85％

[0077] Embodiment 8:

[0078] The persistence of antibacterial activity of Klebsiella pneumoniae phage GRNKpP10

[0079] A single colony of Klebsiella pneumoniae KP002 was picked from the plate, inoculated into 3 mL of TSB medium, and cultured at 37°C and 150 rpm for 8 h to obtain a host bacterial solution for later use. This example investigates the antibacterial persistence of Klebsiella pneumoniae phage GRNKpP10, and the experimental setup is as follows.

[0080] (1) Negative control group: only bacteriophage GRNKpP10 was added at a concentration of 1×10 6 pfu / mL.

[0081] (2) Positive control group: only KP002 was added, and the inoculation amount was 1%.

[0082] (3) KP002+GRNKpP10 group: bacteriophage was added at a concentration of 1×106 pfu / mL; KP002 was added at an inoculum amount of 1%.

[0083] The experiment lasted for 12 hours, and the OD of each group was measured at 1 hour, 2 hours, 4 hours, 6 hours, 8 hours, 12 hours, and 24 hours. 600 .

[0084] The experimental results show that the OD of KP002+GRNKpP10 group was 1h-24h 600 All were maintained below 0.1, and the OD 600 At 8h OD 600 It reached a maximum value of 2.2, indicating that GRNKpP10 has persistent antibacterial activity and can provide an effective biological measure for the prevention and treatment of Klebsiella pneumoniae in the future.

[0085] The basic principles and main features of the present invention and the advantages of the present invention are shown and described above. For those skilled in the art, it is obvious that the present invention is not limited to the details of the above exemplary embodiments, and the present invention can be implemented in other specific forms without departing from the spirit or basic characteristics of the present invention.

[0086] In addition, it should be understood that although the present specification is described according to implementation modes, not every implementation mode contains only one independent technical solution. This description of the specification is only for the sake of clarity. Those skilled in the art should regard the specification as a whole. The technical solutions in each embodiment may also be appropriately combined to form other implementation modes that can be understood by those skilled in the art.

Claims

1. A Klebsiella pneumoniae phage that can lyse multiple K-capsule Klebsiella pneumoniae and has a long-lasting antibacterial effect ( Klebsiella pneumoniae phage) GRNKpP10, characterized in that: The phage was sent to the China Center for Type Culture Collection for preservation on March 4, 2024. The preservation address is: Wuhan University, Wuhan, China; the preservation number is CCTCC NO: M2024373.

2. A use of the Klebsiella pneumoniae phage GRNKpP10 as claimed in claim 1, characterized in that: It is used in environmental disinfection, animal husbandry, poultry farming, aquaculture or biomedical industry to prepare antibacterial agents for Klebsiella pneumoniae.

3. The use of Klebsiella pneumoniae phage GRNKpP10 according to claim 1, characterized in that: The invention is used for preparing medicines for treating or preventing diseases caused by Klebsiella pneumoniae infection.

4. The use of Klebsiella pneumoniae phage GRNKpP10 according to claim 1, characterized in that: The invention is used for preparing feed or feed additive for antagonizing Klebsiella pneumoniae.

5. The use according to any one of claims 2 to 4, characterized in that: The Klebsiella pneumoniae is K1 capsule type, K11 capsule type, K22 capsule type, K24 capsule type, K31 capsule type, K37 capsule type and K64 capsule type Klebsiella pneumoniae.

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