Phage, composition and application

Abstract

The invention relates to the field of microorganisms, and provides a bacteriophage, a composition and application of the bacteriophage, and the preservation number of the bacteriophage is CCTCC M 2024569. The invention discloses a bacteriophage capable of splitting multiple salmonella and escherichia coli O157: H7, and a bacteriophage composition comprising the bacteriophage. The effective components of the bacteriophage composition comprise the bacteriophage or the bacteriophage composition. The bacteriophage provided by the invention has good biological characteristics, does not contain virulence genes and antibiotic resistance genes, and is wide in host spectrum and strong in splitting property. The bacteriophage preparation provided by the invention can be used as a biological prevention and control preparation to control infection of salmonella and escherichia coli O157: H7 antibiotic-resistant strains in food.

Description

Technical Field

[0001] The present invention relates to the field of microbial technology, and more specifically, to a bacteriophage, a composition and an application thereof. Background Art

[0002] Food safety issues caused by foodborne pathogens pose a serious threat to human health. According to WHO estimates, 420,000 people died worldwide in 2020, and huge economic losses were also caused, with an estimated loss of US$110 billion. There are more than 30 types of foodborne pathogens that cause human diseases, of which Escherichia coli O157:H7 and Salmonella are the main foodborne pathogens. Escherichia coli O157:H7 is a dangerous foodborne pathogen with strong resistance, acid and low temperature resistance, Gram-negative staining, and is a common serotype of enterohemorrhagic Escherichia coli. It mainly infects humans through contaminated food and water, and can cause diarrhea, hemorrhagic colitis, hemolytic uremic disease and even death in humans, with a mortality rate of 5-10%. Symptoms of salmonellosis caused by Salmonella infection include nausea, vomiting, diarrhea, fever, abdominal pain, and sometimes bloody stools and even death. There are many serotypes of Salmonella. Currently, 1,500 serotypes that are pathogenic to humans have been identified. Among them, Salmonella Enteritidis and Salmonella Typhimurium are the most harmful, with more than half of the cases being caused by Salmonella Enteritidis and Salmonella Typhimurium.

[0003] Finding and using natural bio-inhibitors to make food safe for human consumption without negatively affecting the taste, texture and nutritional quality of food is an ongoing challenge faced by the food industry around the world, especially as the global population continues to grow and multi-antibiotic resistance in pathogenic bacteria is becoming increasingly common. As a potential prevention and control method, phage biocontrol technology has received a lot of attention in recent years. Bacteriophages have the characteristics of strong specificity, no toxic side effects, and wide distribution. Compared with the physical (steam, high temperature, irradiation) and chemical (bactericides and preservatives) bactericidal or bacteriostatic methods used in traditional food production and processing, the advantage of using phage preparations is that they will not affect the appearance and flavor of food. However, the activity of phages can also be affected by environmental conditions such as temperature and pH, which limits their practical application.

[0004] At present, there are related patent technologies used in food, such as Zhou Yang et al. (authorization announcement number: CN 109825479B) disclosed a wide-spectrum Salmonella phage LPSTLL and its application. The Salmonella phage LPSTLL can lyse 13 serotypes of Salmonella such as Typhimurium, Enteritis, Dublin, and multiple strains of Salmonella with drug resistance. However, this patent technology only lyses Salmonella, and cannot control Escherichia coli O157:H7. Wu Guoping et al. (application publication number: CN 114196637 A) disclosed a Salmonella phage (salmonella sp.phage) JNwz02 and its application. The Salmonella phage JNwz02 can lyse 8 serotypes of Salmonella such as Stanleyville, Hillingdon, and Typhoid, and can also lyse enterohemorrhagic Escherichia coli O157:H7. The phage can inhibit Stanleyville Salmonella and Enterohemorrhagic Escherichia coli O157:H7 in braised duck neck. However, the patent does not describe the effect of removing biofilm.

[0005] Although some relevant patents in China have disclosed the preparation of bacteriophages or phage preparations and their use in food, due to the diversity of pathogenic bacteria serotypes and the host specificity of phages, and on the other hand due to the increasingly serious bacterial resistance, there is an urgent need to screen for dominant phages with strong tolerance, a wide host range, and the ability to lyse resistant bacteria, and to establish and use large-capacity and diverse phage reserves to meet the demand.

[0006] To this end, we propose a bacteriophage and a composition and application to solve the above problems. Summary of the invention

[0007] Technical issues to be solved

[0008] The present invention overcomes the shortcomings of the prior art and provides a broad-spectrum and highly effective natural biological control agent and a preparation method and application thereof, which can be used for ready-to-eat food or fresh food to prevent and control the contamination of Escherichia coli O157:H7 and Salmonella.

[0009] Technical Solution

[0010] To solve the above problems, the present invention adopts the following technical solutions.

[0011] A bacteriophage, the deposit number of the bacteriophage is: CCTCC M 2024569.

[0012] Such as the application of bacteriophage in instant food or fresh food.

[0013] A phage composition comprises the above-mentioned phage.

[0014] A phage preparation, the active ingredient of which includes the above-mentioned phage or the above-mentioned phage combination.

[0015] A method for preparing phage, comprising:

[0016] Take a water sample, add a calcium chloride solution with a concentration of 0.2 mol / L, mix well and let stand for 2 h;

[0017] Filter through a 0.22 μm filter membrane;

[0018] Take 50 mL of the filtrate, add 50 mL of 2×LB liquid medium, and then add 2.5 mL of Salmonella bacteria liquid in the logarithmic phase. Place it on a shaker and culture overnight at 37 °C and 180 r / min;

[0019] The next day, centrifuge the culture solution at 10 °C and 3000 g / min for 10 min;

[0020] Take the supernatant and filter it through a 0.22 μm filter membrane to obtain the phage stock solution;

[0021] Use SM buffer to dilute the phage stock solution by 10-fold gradient. Take 100 μL of the diluted solution and mix it with 100 μL of Salmonella bacteria in the logarithmic phase. Incubate for 15 min, add 8 mL of molten 0.75% agar semi-solid LB medium, mix well, pour the total volume onto an LB plate containing 1.5% agar, and incubate at 37 °C for 18 - 24 h;

[0022] Take the clear phage plaque into 5 mL of LB medium, and then add 100 μL of the host bacteria in the logarithmic phase. Culture overnight at 37 °C and 180 rpm with shaking.

[0023] A method for preparing a phage preparation, inoculating the above-mentioned phage stock solution and the host bacteria in the logarithmic growth phase at a ratio of 1:100 into 5 L of LB liquid medium for scale-up culture for 6 h, at a temperature of 37 °C and a stirring speed of 100 rpm / min;

[0024] Coarse filtration to remove large particle impurities to obtain the initial phage filtrate;

[0025] Use the ultrafiltration method to remove small molecule substances such as endotoxin in the filtrate;

[0026] Use 5 L of eluent to elute the phage, and the phage titer in the eluent is 109 - 1011 PFU / mL.

[0027] Preferably, the coarse filtration uses a filter membrane with a diameter of 0.45 μm.

[0028] Preferably, the ultrafiltration method uses an ultrafiltration membrane with a molecular weight cut-off of 100 - 200 kDa.

[0029] Preferably, the solvent of the eluent is pure water, and the solutes are protocatechuic acid protectant and glycerol protectant.

[0030] Preferably, the ratio of the protocatechuic acid protective agent is 0.03-0.05%, and the ratio of the glycerol protective agent is 0.1-0.4%.

[0031] Beneficial effects: Compared with the prior art, the advantages of the present invention are:

[0032] 1. The bacteriophage provided by the present invention is a new type of multivalent bacteriophage with good biological properties and no virulence genes or antibiotic resistance genes. The bacteriophage preparation provided by the present invention can be used as a biological control preparation to control the infection of Salmonella and Escherichia coli O157:H7 antibiotic-resistant strains in food. BRIEF DESCRIPTION OF THE DRAWINGS

[0033] Figure 1 It is in the form of bacteriophage.

[0034] Figure 2 Temperature tolerance of bacteriophage.

[0035] Figure 3 pH tolerance of the phage.

[0036] Figure 4 Circle diagram of the complete phage genome.

[0037] Figure 5 This is a phylogenetic tree based on the amino acid sequence of the large subunit of bacteriophage terminase.

[0038] Figure 6 A clear picture of the effect of bacteriophage on Salmonella biofilm. DETAILED DESCRIPTION

[0039] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention; it is obvious that the described embodiments are only part of the embodiments of the present invention, rather than all the embodiments, and all other embodiments obtained by ordinary technicians in this field based on the embodiments of the present invention without making creative work are within the scope of protection of the present invention.

[0040] The embodiments of the present application provide a bacteriophage, a composition and an application, which overcome the shortcomings of the prior art and provide a broad-spectrum and highly effective natural biological control agent and a preparation method and application thereof, which can be used for ready-to-eat food or fresh food to prevent and control the contamination of Escherichia coli O157:H7 and Salmonella, and can lyse a variety of Salmonella and Escherichia coli O157:H7. The culture mentioned in this application has been deposited with the China Center for Type Culture Collection on March 27, 2024, with the deposit number of the collection center: CCTCCNO: 2024569. The name of the culture requested for deposit and the indicated identification characteristics: Salmonella phage SF02, Salmonella phage SF02. The culture requested for deposit is accompanied by a scientific description. Deposit address: Wuhan University, Wuhan, China.

[0041] The technical solution in the embodiment of the present application is to solve the above technical problems, and the overall idea is as follows:

[0042] The present invention uses a polyvalent virulent bacteriophage isolated from river water as an antibacterial agent, confirms its safety and effectiveness through biological property research and whole genome analysis, and then prepares a high-titer phage preparation, which is applied to various foods to test the control effect on bacteria.

[0043] Reference Figure 1-Figure 6 , the present invention comprises the following steps:

[0044] 1. Isolation and purification of bacteriophage

[0045] Take 200 mL of water sample, add calcium chloride solution with a final concentration of 0.2 mol / L, mix thoroughly and let stand for 2 hours, then filter with a 0.22 μm filter membrane, take 50 mL of filtrate and add 50 mL of 2×LB liquid culture medium, then add 2.5 mL of logarithmic phase Salmonella liquid, place on a shaker, and culture overnight at 37°C and 180 r / min. The next day, centrifuge the culture at 10°C and 3000 g / min for 10 minutes. Take the supernatant and filter it through a 0.22 μm filter membrane to obtain the phage stock solution.

[0046] Then, the double-layer plate method was used to isolate the phage. The phage stock solution was diluted 10 times with SM buffer, and 100 μL of the dilution was mixed with 100 μL of logarithmic phase Salmonella, incubated for 15 minutes, and 8 mL of molten 0.75% agar semi-solid LB medium was added, mixed, and the total volume was poured onto an LB plate containing 1.5% agar, and incubated at 37°C for 18-24 hours (double-layer agar plate method). The transparent plaque was placed in 5 mL of LB medium, and 100 μL of logarithmic phase host bacteria was added, and cultured overnight at 37°C and 180 rpm. The double-layer plate method was repeated 3 times until the plaque morphology was consistent.

[0047] 2. Preparation of phage preparations

[0048] The phage stock solution and the logarithmic growth phase host bacteria were inoculated in a 5L LB liquid culture medium at a ratio of 1:100 for expansion culture for 6 hours at a temperature of 37°C and a stirring speed of 100rpm / min, followed by coarse filtration (filter membrane diameter of 0.45μm) to remove large particle impurities to obtain the phage primary filtrate, and then ultrafiltration (ultrafiltration membrane molecular weight cutoff of 100-200kDa) was used to remove small molecules such as endotoxins in the filtrate, and then 5L elution liquid was used to elute the phage, and the phage titer in the eluate was 109-1011PFU / mL. The solvent of the eluate was pure water, and the solute was a protocatechuic acid protective agent (0.03-0.05%) and a glycerol protective agent (0.1-0.4%).

[0049] 3. Storage of phage preparations

[0050] The prepared phage preparation is packaged and stored in a dry, cool, dark place.

[0051] See also Figure 1-Figure 6 , a bacteriophage, a composition and an application.

[0052] Specific Example 1 Observation of bacteriophage microscopic morphology

[0053] Take 10 μL of purified phage (109 PFU / mL) and drop it on the front of the copper mesh. After drying at room temperature for 5 minutes, stain it with 1% phosphotungstic acid for 1-2 minutes, dry it at room temperature, and observe the phage morphology using a JEM-1400FLASH transmission electron microscope. The phage has an inelastic tail and a polyhedral head. The tail is about 220 nm long and the head diameter is about 65 nm ( Figure 1 ).

[0054] Specific Example 2 Phage tolerance analysis

[0055] 1mL of phage suspension (109PFU / mL) was incubated at 30, 40, 50, 60, 70, and 80℃ for 1h each, and 100μL of phage suspension was collected and 10-fold gradient dilution was performed to determine the phage titer. 100μL of phage suspension was added to 900μL of SM buffer with different pH values, incubated at 37℃ for 1h, and then the phage titer was determined using the double-layer plate method. The experiment was repeated 3 times. The results are shown in Figure 2. Figure 2 As shown, the phage remained stable in the temperature range of 30℃ to 60℃, with a titer of more than 109PFU / mL. After being treated at 70℃ for 1 hour, the titer remained at 5Log10 PFU / mL, which has a high titer. After being treated at pH3-11 for 1 hour, the titer was stable and remained above 109PFU / mL. Phages still survived after being treated at pH2 and pH12 for 1 hour, which has a wide temperature and pH tolerance range.

[0056] Specific Example 3: Phage Host Spectrum Test and Host Drug Resistance Analysis

[0057] According to the CLSI standard, the sensitivity of all test strains to 11 antibiotics such as ampicillin was determined by the disk diffusion method. Then, the lytic activity against 42 Salmonella strains and 2 Escherichia coli strains was determined by the spot test. 100 μL of the bacterial culture solution in the logarithmic phase was spread on an LB agar plate. Then, 10 μL of the phage (109 PFU / mL) was spotted on each bacterial plate, and 10 μL of SM buffer was used as a control. After incubating overnight at 37 °C in a constant temperature incubator, the transparency of the spotted area was observed. The results are shown in Table 1. The phage had lytic activity against 33 Salmonella strains and 1 Escherichia coli strain, and the lysis rate was 77.3%. Among the 33 Salmonella strains that could be lysed, 22 Salmonella strains had multidrug resistance (resistant to 3 or more antibiotics). The results showed that it could cross-species lyse Salmonella and Escherichia coli, including multidrug-resistant strains.

[0058] Table 1 Antibiotic Resistance of Different Strains and Host Range of Isolated Phages

[0059]

[0060]

[0061]

[0062]

[0063] Note: AMP: Ampicillin; AMC: Amoxicillin; CTX: Cefotaxime; TET: Tetracycline; GEN: Gentamicin; NEO: Neomycin; FFC: Florfenicol; CIP: Ciprofloxacin; NAL: Nalidixic acid; SXT: Co-trimoxazole; C: Chloramphenicol.

[0064] "+++" The plaque edge is clear, transparent, "++" The phage plaque is clearer and transparent, "+" The plaque is turbid, "-" There is no plaque.

[0065] Specific Example 4: Phage Whole Genome Sequencing

[0066] The whole genome of the phage was sequenced by an Illumina sequencer, and then assembled and compared for annotation. According to the annotation results, it was found that the genome did not contain virulence genes or antibiotic resistance genes. The full length of the genome was 158,384 bp, the GC content was 49.98%, and it contained 209 open reading frames ( Figure 3), 52 of which have significant homology with known functional genes. The similarity with other phages was analyzed by BLAST-N, and the results showed that the genome had the highest similarity with the genome of Escherichia coli phage CBA120 (accession number: NC016570.1) of the Kuttervirus genus of the Ackermannviridae family, with a query coverage of 92% and an identification coverage of 98.85%. The Mauve software was used to perform a colinearity analysis on phage CBA120, phage BSP101 and phage SWJM-01 with high similarity to the whole genome, and it was found that a large number of rearrangements and ectopic positions occurred in the genome, indicating that it is a new phage. The amino acid sequence of the large subunit of the terminase that was conserved during evolution was selected.

[0067]

[0068] Phylogenetic tree was constructed with phage sequences similar to those in NCB I. The results are shown in the figure. The phage has a close evolutionary relationship with Kutterv i rus virus genus Escherichia coli phage vB EcoA 4HA11 and Salmonella phage SenASZ3, with the highest homology. In summary, it is a new type of Kutterv i rus virus genus polyvalent phage.

[0069] Specific Example 5 The lysis effect of bacteriophage preparation on bacteria in chicken feet:

[0070] Fresh ready-to-eat chicken feet were purchased from the supermarket, and then each portion of about 25g ready-to-eat chicken feet was placed in a water bath at 100℃ for 20 minutes to kill most of the initial bacteria. Then, the sample was immersed in a culture solution of Salmonella or Escherichia coli O157:H7 diluted to 104CFU / mL for 10 minutes, and then placed on a clean bench to air dry for 30 minutes to allow the bacteria to fully adsorb on the sample surface. Then 500μL of 5×109PFU / mL phage preparation was added to the sample surface to make the concentration 107PFU / g. After treatment, the sample was placed at 4℃ and homogenized with 75mL eluent for 30s at 0, 2, 4, 6, 8, and 10h. Centrifuge at 3000g for 10min to remove large particles. The supernatant was then diluted 10 times in a gradient and counted with a differential medium at an appropriate dilution. Each experiment was repeated three times.

[0071] As shown in Table 2, during the storage at 4°C for 10 hours, the number of bacteria on the surface of the chicken feet treated in Comparative Example 1 remained stable, the number of Salmonella was maintained at more than 4.18Log10 CFU / g, and the number of Escherichia coli O157:H7 was maintained at more than 4.15Log10 CFU / g. However, after treatment with the phage preparation, the number of bacteria decreased significantly (P<0.05), the number of live Salmonella rapidly decreased to 3.89Log10 CFU / g at 0h, and no Salmonella was detected after 2h of treatment. In addition, the number of Escherichia coli O157:H7 also decreased rapidly, from 3.67Log10 CFU / g at the beginning to undetectable after 2h.

[0072] Table 2 Inhibitory effect of Example 5 and Comparative Example 1 on Salmonella or Escherichia coli O157:H7 in ready-to-eat chicken feet

[0073]

[0074]

[0075] Note: “ / ” indicates no Salmonella or E. coli O157:H7 was detected.

[0076] Specific Example 6: Lysis effect of bacteriophage preparation on bacteria in pork

[0077] Fresh pork was cut into 2×2×1cm3 sizes and sterilized with ultraviolet light for 30 minutes. The pork was then artificially contaminated with Salmonella and Escherichia coli O157:H7, so that the final bacterial concentration on the meat surface (2×2cm2) was 104CFU / cm2, and dried at room temperature for 30 minutes. It was then treated with 200μL of 2×109PFU / mL of phage preparation, and the sample was then placed at 4°C. Samples were taken at 0, 2, 4, 6, 8, and 10 hours, and the eluent was used for oscillation elution. The colony counts of Salmonella and Escherichia coli O157:H7 in the samples were determined using identification medium, and each experiment was repeated three times.

[0078] In comparative example 2, the bacterial count in pork did not decrease, while in example 6, the counts of Salmonella and E. coli O157:H7 decreased from 4.21Log10 CFU / cm2 and 4.22Log10 CFU / cm2 at the beginning to 3.54Log10 CFU / cm2 and 3.74Log10 CFU / cm2, respectively, and the target bacteria were detected after 2 hours of treatment. This is because the phage quickly lyses the host bacteria after contact with the bacteria. In general, the phage has a significant antibacterial effect on Salmonella or E. coli O157:H7 in ready-to-eat chicken feet and pork, indicating that the phage preparation has the ability to be used in ready-to-eat meat foods to prevent and control contamination by Salmonella and E. coli O157:H7.

[0079] Table 3 Inhibitory effects of Example 6 and Comparative Example 2 on Salmonella or Escherichia coli O157:H7 in pork

[0080]

[0081]

[0082] Note: “ / ” indicates no Salmonella or E. coli O157:H7 was detected.

[0083] Specific Example 7: The lysis effect of bacteriophage preparation on Salmonella biofilm.

[0084] Salmonella cultured to the logarithmic growth phase was added to a 96-well plate and cultured for 12, 24, 36, and 48 hours, then the excess bacterial solution was slowly aspirated with a pipette, and then gently rinsed 3 times with PBS buffer. Subsequently, 200 μL of phage preparation (108 PFU / mL) was added and treated for 24 hours. After the treatment, the phage preparation was aspirated, rinsed with PBS buffer, and then 200 μL of 95% methanol solution was added, fixed at room temperature for 30 minutes, and then rinsed 3 times with PBS. Finally, an appropriate amount of 1% crystal violet solution was added for staining for 10 minutes, and the absorbance value at 600 nm was measured.

[0085] The results are as follows Figure 6 As shown, the OD600nm value of the Salmonella biofilm gradually increased with the increase of culture time. After being treated with the phage preparation, the OD600nm value was significantly lower than that of Comparative Example 3, indicating that the phage preparation has a significant ability to eliminate biofilms of different growth times.

[0086] Comparative Example 1

[0087] The steps of treating ready-to-eat chicken feet in Comparative Example 1 were similar to those in Example 6, except that 500 μL of phage preparation was replaced with an equal volume of SM buffer for treatment.

[0088] Comparative Example 2

[0089] The steps of processing pork in Comparative Example 2 were similar to those in Example 7, except that 200 μL of phage preparation was replaced with an equal volume of SM buffer for processing.

[0090] Comparative Example 3

[0091] The steps of treating the Salmonella biofilm in Comparative Example 3 were similar to those in Example 8, except that the phage preparation treatment was replaced by an equal amount of SM buffer treatment.

[0092] Working principle:

[0093] 1. This phage has the ability to lyse Escherichia coli and Salmonella across species. According to the results of whole genome sequencing and annotation, it has four tail fiber proteins, which can bind to Escherichia coli antigens or Salmonella-specific receptors respectively, thereby exerting a cross-species lysis effect, so it has a wide host range.

[0094] 1. The bacteriophage provided by the present invention can lyse a variety of Salmonella and Escherichia coli O157:H7, and the lysed bacteria are mostly antibiotic-resistant strains or multi-drug-resistant strains, providing a potential backup phage to cope with the increasingly serious bacterial resistance crisis.

[0095] 2. The complete gene of the bacteriophage provided by the present invention does not contain virulence genes, antibiotic resistance genes, etc. The bacteriophage of the present invention is a phage of natural origin.

[0096] 3. The titer of the bacteriophage provided by the present invention is maintained at 5Log10 PFU / mL after being treated at 70°C for 1 hour, which has a high titer. The titer is stable and remains above 109 PFU / mL after being treated at pH 3-11 for 1 hour. The bacteriophage still survives after being treated at pH 2 and pH 12 for 1 hour, which has a wide temperature and pH tolerance range.

[0097] 4. The bacteriophage provided by the present invention has a good lysis and inhibition effect on bacteria in a liquid environment. When the MOI is greater than or equal to 10, the production of bacteria can be completely inhibited.

[0098] 5. The bacteriophage preparation provided by the present invention has a strong lysis effect on the bacteria on the surface of ready-to-eat chicken feet and fresh pork. After only 2 hours of treatment, all the bacteria on the surface of ready-to-eat chicken feet and fresh pork were removed.

[0099] 6. The bacteriophage preparation provided by the present invention has a good effect in removing Salmonella biofilm. The bacterial biofilm can improve the tolerance of bacteria to adverse external environments, thereby improving their survival ability. In addition, the biofilm can also reduce the sensitivity of bacteria to antibiotics, thereby increasing drug resistance.

[0100] The above is only a preferred specific implementation of the present invention; however, the protection scope of the present invention is not limited thereto. Any technician familiar with the technical field can make equivalent replacements or changes according to the technical solution and its improved conception within the technical scope disclosed by the present invention, which should be covered by the protection scope of the present invention.

[0101] It should be noted that, in this article, relational terms such as first and second, etc. are only used to distinguish one entity or operation from another entity or operation, and do not necessarily require or imply any such actual relationship or order between these entities or operations. Moreover, the terms "include", "comprise" or any other variants thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, the elements defined by the sentence "comprise a ..." do not exclude the existence of other identical elements in the process, method, article or device including the elements.

[0102] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features thereof may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A bacteriophage, characterized in that: The deposit number of the bacteriophage is: CCTCC M 2024569.

2. Use of the bacteriophage as claimed in claim 1 in instant food or fresh food.

3. A bacteriophage composition, characterized in that: Comprising the bacteriophage as claimed in claim 1.

4. A bacteriophage preparation, characterized in that: The active ingredient comprises the bacteriophage according to claim 1 or the bacteriophage composition according to claim 3.

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