Phage vB_VneM_XG7 and application thereof

Abstract

The application provides a bacteriophage vB_VneM_XG7 and an application thereof, and belongs to the technical field of microorganisms. The bacteriophage vB_VneM_XG7 is preserved in the China General Microbiological Culture Collection Center, and the preservation number is CGMCC No. 46367. The bacteriophage vB_VneM_XG7 provided by the application has strong tolerance to temperature and pH, can adapt to various environments, does not contain virulence genes and antibiotic resistance genes in the genome of the bacteriophage vB_VneM_XG7, has good biological safety, has the ability of efficiently lysing pathogenic vibrio, can effectively target the disease problems caused by pathogenic vibrio, has great advantages in treating aquaculture bacterial diseases, will not cause bacterial drug resistance and resistance, will not cause secondary pollution to the breeding environment, and is green and environment-friendly.

Description

Technical Field

[0001] This invention belongs to the field of microbial technology, and in particular relates to a bacteriophage vB_VneM_XG7 and its applications. Background Technology

[0002] Large-scale aquaculture, such as fish, scallops, and shrimp, is often affected by pathogens. These bacteria can indirectly affect aquatic crops through direct contact or by secreting toxins, leading to disastrous consequences for the aquaculture industry and the ecological balance of aquaculture areas.

[0003] Currently, various pathogenic bacteria have been isolated and reported, mainly including Vibrio ( Vibrio ), Pseudomonas ( Pseudomonas Aeromonas hydrophila ( ) Aeromonas hydrophila Edwardsiella tarda ( ), Edwardsiella tarda ( Edwardsiella tarda Streptococcus ( Streptococcus (etc.). Among them, Vibrio is one of the most common diseases in marine farmed animals.

[0004] Vibrio ( Vibrio ) belongs to the phylum Proteobacteria ( Proteobacteria ), Gamma-Proteobacteria ( γ- Proteobacteria ), Vibriales ( Vibrionales Vibrioceae ( Vibrionaceae These bacteria are typically arc-shaped or rod-shaped, possess polar flagella, and are Gram-negative. They are a type of heterotrophic bacteria widely found in marine and freshwater environments. Extensive evidence indicates that this genus not only possesses rich metabolic capabilities, effectively utilizing various carbon sources to synthesize complex compounds and gaining an advantage in competition with other microorganisms, but also secretes a variety of extracellular active substances, such as extracellular enzymes and toxins. It has been confirmed that many Vibrio bacteria produce extracellular enzymes and toxins with strong pathogenic activity, capable of efficiently infecting and killing host animal cells, such as *Vibrio splenita*. Vibrio splendidus It has been proven to cause mass mortality in scallop larvae; in addition, Vibrio thaliana... Vibrio mediterranei It has been identified as the pathogen causing mass mortality in razor clam larvae, and its secreted extracellular products are highly toxic to the larvae; *Vibrio neocalidonia*. Vibrio neocaledonicus Can cause whiteleg shrimp ( Litopenaeus vannameiAn outbreak of "Shrimp Postlarva Bacterial Vitrified Syndrome" (BVS) has occurred at a hatchery. This bacterium is highly pathogenic to PL5 and PL20 larvae and juvenile shrimp of Litopenaeus vannamei, causing infected shrimp larvae to exhibit weak vitality, dark and cloudy body color, blurred hepatopancreas outline, pale yellow discoloration, tissue disintegration with a hyalinized appearance, empty intestines and stomachs, and ultimately acute death. Furthermore, a large number of Vibrio bacteria have developed significant resistance to commonly used antibiotics (such as rifampin, pipemidic acid, streptomycin, polymyxin, penicillin, and trimethoprim-sulfamethoxazole). Therefore, the pathogenic Vibrio bacteria seriously affect the development of the aquaculture industry.

[0005] Currently, while the rapid development of scallop and other shellfish farming, as well as shrimp farming, has brought considerable economic benefits, disease problems have gradually become a major bottleneck restricting the healthy development of aquaculture. Large-scale diseases can even lead to the death of more than half of the aquatic animals in a farm. Research on diseases in large-scale economic aquatic animals is mainly limited to visual observation of symptoms, microscopic observation, and morphological description of pathogens. Although many studies have confirmed that mass mortality in aquatic animals is related to bacterial diseases, the pathogenic causes and mechanisms of these pathogens, as well as the disease resistance and defense systems of aquatic animals, remain unclear. Traditional treatment methods using antibiotics and chemical disinfectants have generated numerous problems, such as bacterial resistance, secondary pollution of the aquaculture environment, safety issues caused by pathogen transmission, and imbalance of the aquatic micro-ecology. Therefore, there are no completely effective preventive measures for diseases in large-scale economic aquatic animals. The current common practice is to reduce diseases by strengthening management and controlling environmental factors (such as temperature, salinity, and water quality) during the farming process, but this is not conducive to larger-scale farming. Meanwhile, because pathogenic Vibrio bacteria are resistant to commonly used antibiotics, traditional antibiotic treatments have limited effectiveness. This further highlights the importance of finding new prevention and control methods.

[0006] Bacteriophages, as a novel microecological antibacterial agent, have the potential to replace antibiotics in treating diseases due to their environmental friendliness and high specificity. However, bacterial infections in fish, shellfish, and crustaceans in aquaculture are diverse, and there is an urgent need to develop a bacteriophage that can efficiently lyse and infect pathogenic Vibrio bacteria. This is of great significance for the prevention and control of bacterial infections in fish, shellfish, and crustaceans in aquaculture. Summary of the Invention

[0007] Therefore, the purpose of this invention is to provide a bacteriophage vB_VneM_XG7 and its application, wherein the bacteriophage vB_VneM_XG7 has a lytic effect on pathogenic Vibrio.

[0008] To achieve the above-mentioned objectives, the present invention provides the following technical solution: This invention provides a bacteriophage vB_VneM_XG7, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 46367.

[0009] Preferably, the phage vB_VneM_XG7 has a latency period of 20 min and a lysis period of 100 min.

[0010] Preferably, the bacteriophage vB_VneM_XG7 can maintain relatively stable activity between pH 6 and 11 and between -20°C and 70°C.

[0011] The present invention provides a microbial preparation comprising the above-mentioned bacteriophage vB_VneM_XG7.

[0012] The present invention provides a bactericide comprising the bacteriophage vB_VneM_XG7 described above.

[0013] This invention provides a bacteriophage drug formulation, wherein the active ingredient of the bacteriophage drug formulation includes the aforementioned bacteriophage vB_VneM_XG7.

[0014] This invention provides the application of the above-mentioned bacteriophage vB_VneM_XG7, microbial preparations, bactericides, or bacteriophage drug preparations in the preparation of products that kill pathogenic Vibrio.

[0015] This invention provides the application of the above-mentioned bacteriophage vB_VneM_XG7, microbial preparations, bactericides, or bacteriophage drug preparations in the preparation of products for treating pathogenic vibriosis diseases.

[0016] Preferably, the pathogenic Vibrio includes Vibrio neocalidonia.

[0017] Preferably, the phage vB_VneM_XG7 has a lysis rate of 241 PFU / cell against pathogenic Vibrio.

[0018] Compared with the prior art, the present invention has the following beneficial effects: This invention provides a bacteriophage vB_VneM_XG7 and its applications. The bacteriophage vB_VneM_XG7 provided by this invention has strong tolerance to temperature and pH and can adapt to various environments. The genome of bacteriophage vB_VneM_XG7 does not contain virulence genes or antibiotic resistance genes, and has good biosafety. It also has the ability to efficiently lyse pathogenic Vibrio, and can effectively target diseases caused by pathogenic Vibrio. It has great advantages in treating bacterial diseases in aquaculture, specifically: (1) It does not destroy the normal epiphytic flora of seaweed, but only targets the corresponding pathogens, and has strong specificity; (2) It will not cause bacterial drug resistance and resistance; (3) It can rely on the host bacteria to proliferate, and a single dose can achieve the effect of multiple doses of other antibacterial drugs; (4) It has a short residence time in the body and rapid metabolism; (5) It has a short research and development and production time; (6) It will not cause secondary pollution to the aquaculture environment and is green and environmentally friendly.

[0019] Biological Preservation Information: The bacteriophage vB_VneM_XG7 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 46367, deposit date January 6, 2025, at Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, 100101, China. It is classified as *Vibroneocaledonicus phage* and named vB_VneM_XG7. Attached Figure Description

[0020] Figure 1 This is a transmission electron microscope (TEM) image of bacteriophage vB_VneM_XG7. Figure 2 This is a one-step growth curve of bacteriophage vB_VneM_XG7; Figure 3 pH tolerance curves for bacteriophage vB_VneM_XG7; Figure 4 Temperature tolerance curves for bacteriophage vB_VneM_XG7; Figure 5 This is the complete genome map of bacteriophage vB_VneM_XG7. Detailed Implementation

[0021] This invention provides a bacteriophage vB_VneM_XG7, which is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 46367.

[0022] In this invention, the bacteriophage vB_VneM_XG7 is a short-tailed bacteriophage capable of specifically lysing Vibrio spp. The incubation period of vB_VneM_XG7 is 20 min, and the lysis period is 100 min. vB_VneM_XG7 maintains relative stability in activity between pH 6 and 11 and between -20°C and 70°C, making it a virulent bacteriophage adaptable to various environments. The full-length genome of vB_VneM_XG7 is 194309 bp, containing 230 open reading frames (ORFs), of which 135 ORFs (60%) are annotated as putative proteins, and the remaining 95 ORFs (40%) encode functional proteins. Furthermore, the vB_VneM_XG7 genome does not contain virulence genes or antibiotic resistance genes, exhibiting good biocompatibility.

[0023] The bacteriophage of the present invention can be used to lyse pathogenic Vibrio aquatic bacteria, specifically Vibrio neocalidonia. Vibrio neocaledonicus, Therefore, the bacteriophage vB_VneM_XG7 can be used to treat diseases in farmed shrimp caused by pathogenic Vibrio aquaticis.

[0024] The bacteriophage vB_VneM_XG7 provided by this invention is a biological method that can efficiently kill major pathogens in cultured algae. It is highly efficient and environmentally friendly, and can achieve the prevention and control of diseases in cultured algae without disrupting the microecological balance. This is unmatched by antibiotics and other antibacterial drugs.

[0025] The method for culturing bacteriophage vB_VneM_XG7 of the present invention includes: culturing bacteriophage vB_VneM_XG7 and host bacteria at 28°C with shaking to allow the bacteriophage to proliferate.

[0026] The present invention provides a microbial preparation comprising the above-mentioned bacteriophage vB_VneM_XG7.

[0027] In this invention, the microbial preparation further includes pharmaceutically, food, or feed-acceptable excipients.

[0028] The present invention provides a bactericide comprising the bacteriophage vB_VneM_XG7 described above.

[0029] In this invention, the microbial preparation further includes pharmaceutically, food, or feed-acceptable excipients.

[0030] This invention provides a bacteriophage drug formulation, wherein the active ingredient of the bacteriophage drug formulation includes the aforementioned bacteriophage vB_VneM_XG7.

[0031] In this invention, the bacteriophage drug formulation further includes the aforementioned pharmaceutically acceptable excipients. The dosage form of the bacteriophage drug formulation includes granules, injections, or sprays. The term "active ingredient" refers to the main component responsible for treating, alleviating, or preventing a target effect or disease. The active ingredient of this invention includes bacteriophage vB_VneM_XG7, which is capable of killing pathogenic Vibrio or preventing diseases caused by pathogenic Vibrio. Furthermore, the active ingredient of this invention can also be used with other formulations capable of killing bacteria or preventing diseases caused by bacteria. The active ingredient of this invention can also use bacteriophage vB_VneM_XG7 as the sole active ingredient for killing pathogenic Vibrio or preventing diseases caused by pathogenic Vibrio.

[0032] This invention provides the application of the above-mentioned bacteriophage vB_VneM_XG7, microbial preparations, bactericides, or bacteriophage drug preparations in the preparation of products that kill pathogenic Vibrio.

[0033] This invention provides the application of the above-mentioned bacteriophage vB_VneM_XG7, microbial preparations, bactericides, or bacteriophage drug preparations in the preparation of products for treating pathogenic vibriosis diseases.

[0034] In this invention, the product includes a drug. The pathogenic Vibrio includes Vibrio neocalidonia. The bacteriophage vB_VneM_XG7 has a lysis rate of 241 PFU / cell against pathogenic Vibrio.

[0035] In this invention, unless otherwise specified, all raw material components are commercially available products well known to those skilled in the art.

[0036] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0037] Example 1 A method for isolating bacteriophage vB_VneM_XG7, comprising the following steps: (1) Seawater samples were collected from the nearshore area of ​​Qingdao (36°4'22''N, 120°18'33''E), filtered through a 0.22 μm filter membrane, and then serially diluted with SM buffer (0, 10). -1 10 -2 10 -3 10 -4 10 -5 10 -6 10 -7 10 -8 10 -9 10 -10 ), followed by the logarithmic growth phase of Vibrio neocalidonia Vibrio neocaledonicusTake 200 μL of each bacterial culture (host culture) and mix them thoroughly. After incubation for 15 min, culture them using the double-layer plate method.

[0038] (2) After observing the appearance of transparent patches on the plate, use a sterile pipette tip (1 mL) to puncture the entire piece of agar containing the patches and pipette it into 2 mL of SM buffer. Incubate overnight at 4°C. The next day, filter the sample using a sterile filter membrane with a pore size of 0.22 μm to remove agar fragments and Vibrio. Vibrio neocaledonicus After mixed bacterial suspension infection, the double-layer plate method is used for plate pouring. (3) Repeat step (2) 4 times until a single pure phage vB_VneM_XG7 sample is obtained.

[0039] (4) 200 μL of pure bacteriophage vB_VneM_XG7 sample was added to 20 mL of host bacterial culture in the logarithmic growth phase. After culturing in LB medium for 48 h, a mixed bacterial culture was obtained. The mixed bacterial culture was centrifuged at 8000×g, and the liquid in the upper layer was filtered through a 0.22 μm filter membrane. The filtrate was collected and concentrated to 1 mL by ultrafiltration at 3800×g. The filtrate was then transferred to 200 mL of LB medium and cultured for 48 h. After centrifugation at 3800×g, the culture was concentrated again to obtain a mixed culture system. (5) Repeat step (4) until the final mixed culture system is expanded to 1L. Then, use density gradient centrifugation to obtain pure phage enrichment bands, that is, phage vB_VneM_XG7.

[0040] The bacteriophage vB_VneM_XG7 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 46367, deposited on January 6, 2025, at the Institute of Microbiology, Chinese Academy of Sciences, No. 3, No. 1 Beichen West Road, Chaoyang District, Beijing, 100101, China. It is classified as *Vibroneocaledonicus phage* and named vB_VneM_XG7.

[0041] Example 2 Characterization experiments of bacteriophage vB_VneM_XG7 1. Electron microscopy observation experiment A 20 μL drop of the phage enrichment band obtained in Example 1 was added to a dry copper grid. After waiting 15 min, a 2% phosphotungstic acid (PTA) solution was added, and the process was repeated for another 10 min. The grid was then dried under a lamp and observed under a transmission electron microscope. Morphological identification revealed... Figure 1 As shown.

[0042] Figure 1The results showed that the bacteriophage vB_VneM_XG7 is a short-tailed bacteriophage with a symmetrical icosahedral head and a short, thick, non-contractile tail fiber.

[0043] 2. One-step growth curve experiment The specific steps are as follows: (S1) 1 mL of pure bacteriophage vB_VneM_XG7 (MOI=0.01) obtained in Example 1 was placed into an equal volume of host bacteria Vibrio. Vibrio neocaledonicus The mixture was adsorbed in the liquid at 28°C for 17.5 min to obtain a mixed solution.

[0044] (S2) Take 1 mL of the above mixture, centrifuge at 13000×g for 1 min, discard the supernatant, add 1 mL of Zobell2216E culture medium, briefly vortex to mix and precipitate; (S3) Repeat step (S2) three times to remove unadsorbed phages and obtain the adsorbed mixture.

[0045] (S4) The adsorbed mixture was put back into 50 mL of Zobell 2216E culture medium, and incubated at 28℃ with shaking for 3 hours. During the first 30 minutes, samples were taken every 5 minutes, then every 10 minutes for the next 30 minutes, and then every 30 minutes for the last 2 hours. Each sample was 1 mL and three parallel samples were set up.

[0046] (S5) Place 20 μL of glutaraldehyde into each sample, fix in the dark for 15 min, and immediately freeze with liquid nitrogen; then thaw the sample at room temperature, dilute appropriately with SM buffer (pH 8), and stain the sample with DNA staining agent SYBR Green I at 80℃ for 10 min.

[0047] (5) At the same time, take 1 mL of the host bacterial culture (Vibrio neocaledonicus solution) at the beginning of the timing and put it into 20 μL of glutaraldehyde. Fix it in the dark for 15 min and then freeze it with liquid nitrogen immediately. Thaw the sample at room temperature and dilute it appropriately with TE buffer (Tris-EDTA; pH=8). Stain the sample with DNA staining agent SYBR Green I at 80℃ for 1 h.

[0048] (6) Flow cytometry was used to determine the abundance of bacteriophages and the initial abundance of infected host bacteria, and a one-step growth curve of the bacteriophages was plotted, such as... Figure 2 As shown.

[0049] from Figure 2It can be seen that the incubation period of bacteriophage vB_VneM_XG7 is approximately 20 minutes; then it enters the exponential burst phase (lysis phase), which takes about 100 minutes, with a lysis rate of 241 PFU / cell; indicating that bacteriophage vB_VneM_XG7 is effective against Vibrio. Vibrio neocaledonicu It has strong cleavage ability and infectivity.

[0050] Example 3 Stability test of bacteriophage vB_VneM_XG7 The bacteriophage vB_VneM_XG7 described in this embodiment is the bacteriophage vB_VneM_XG7 prepared in Example 1.

[0051] 1. pH stability Add 100 μL of phage vB_VneM_XG7 sample (10 6 Add 900 μL of buffer solution with different pH values ​​(5, 6, 7, 8, 9, 10, 11, and 12) to each sample (PFU / mL), and incubate at 28°C for 2 h. Take 200 μL of each sample at different pH gradients and add an equal volume of host bacterial culture (Vibrio) to each sample. Vibrio neocaledonicus Mix the liquid and incubate for 15 minutes, then pour into double-layer plates and incubate at 28°C for 12 hours. Calculate and plot the pH effect trend graph, and repeat each pH value three times.

[0052] like Figure 3 The results showed that bacteriophage vB_VneM_XG7 was relatively stable in the pH range of 6 to 11.

[0053] 2. Thermal stability Take 200 μL of phage vB_VneM_XG7 sample (10 6 (PFU / mL; pH=7.0) were incubated at -20℃, 4℃, 25℃, 35℃, 45℃, 55℃, 65℃, and 75℃ for 2 hours; after the temperature in each tube returned to room temperature, 200 μL of host bacterial culture (Vibrio) was added. Vibrio neocaledonicus After 15 minutes of inoculation, the solution was poured into double-layer plates and incubated at 28°C for 12 hours. The number of plaques was then counted, and each temperature was repeated three times.

[0054] like Figure 4 Experimental results show that bacteriophage vB_VneM_XG7 maintains a very stable titer between -20℃ and 45℃, and its viability only begins to decline after 55℃.

[0055] Example 4 Purification and sequencing of the bacteriophage vB_VneM_XG7 genome The specific steps are as follows: (1) The phage vB_VneM_XG7 sample from Example 1 was used to extract and purify the genome sequentially using the HP Viral DNA / RNA Kit (OMEGA, M6246-01) and the DNeasy Power Clean pro Cleanup Kit (Qiagen, 12997-50).

[0056] (2) The purified DNA sample was broken down to 350bp by sonication. Then the DNA fragment ends were repaired, tailed, and folded. After PCR, the product was purified (AMPure-XP system) and a DNA library was constructed. Then, the DNA was sequenced using Illumina NovaSeq PE150.

[0057] (3) The open reading frames (ORFs) in the phage sequence were determined using RAST (https: / / topaz.gatech.edu / GeneMark / ) and GeneMarks (http: / / topaz.gatech.edu / GeneMark / ) software, respectively. Then, the ORFs were compared with the non-redundant (NR) protein database based on the BLASTP algorithm in NCBI (https: / / www.ncbi.nlm.nih.gov / ), the Pfam (https: / / pfam.xfam.org / search / sequence) database, and the PDB (http: / / www.rcsb.org / ) database. The comparison result with the highest score (E-value ≤ 10) was selected. -5 The phage genome was filtered and integrated using id-entity>40% and coverage>40% to obtain the final annotation information.

[0058] The complete genome map of bacteriophage vB_VneM_XG7 is as follows: Figure 5 As shown, the specific sequence information of the whole genome is shown in Table 1.

[0059] Sequencing revealed that the full-length genome of bacteriophage vB_VneM_XG7 is 194,309 bp, containing a total of 230 open reading frames (ORFs).

[0060] Figure 5 The results indicate that the functions of the bacteriophage vB_VneM_XG7 genome include assisting metabolism, DNA replication and metabolism, transcriptional regulation, structure and packaging, and lysis.

[0061] The results in Table 1 indicate that the vB_VneM_XG7 genome does not contain virulence genes or antibiotic resistance genes, demonstrating good biocompatibility. Of the ORFs, 135 (60%) were annotated as putative proteins, while the remaining 95 (40%) encoded functional proteins.

[0062] Table 1. Predicted ORFs for bacteriophage vB_VneM_XG7

[0063] This invention provides a strain capable of efficiently degrading Vibrio neocaledonicus The strain's bacteriophage vB_VneM_XG7. Bacteriophage vB_VneM_XG7 can specifically lyse... Vibrio neocaledonicus This rapidly reduces the number of host cells in the sample within a short period. According to one-step growth curve data, the incubation period of bacteriophage vB_VneM_XG7 is only 20 minutes. After 20 minutes, a large number of bacteriophages complete replication, rapidly lysing the host bacteria and clarifying the previously turbid host bacterial solution. Simultaneously, according to data from pH and temperature tolerance experiments, this bacteriophage can withstand pH values ​​between 6 and 11 and temperatures up to -20°C. o C~70 o The fact that C maintains relative stability of activity indicates that bacteriophage vB_VneM_XG7 can effectively adapt to a variety of harsh environments and achieve highly efficient prevention and control of diseases in aquaculture, especially shrimp farming.

[0064] In summary, bacteriophage vB_VneM_XG7 is a strain capable of efficiently lysing pathogenic algae. Vibrio neocaledonicus These are virulent bacteriophages with excellent adaptability to complex marine environments. For those composed of... Vibrio neocaledonicus The treatment of bacterial vitrification in farmed shrimp has unparalleled advantages over traditional antibiotic treatment, and is of great significance for promoting the development of shrimp farming quality and yield.

[0065] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A bacteriophage vB_VneM_XG7, characterized in that, The bacteriophage vB_VneM_XG7 is deposited at the China General Microbiological Culture Collection Center (CGMCC) with accession number CGMCC No. 46367.

2. The bacteriophage vB_VneM_XG7 according to claim 1, characterized in that, The phage vB_VneM_XG7 has a latency period of 20 min and a lysis period of 100 min.

3. The bacteriophage vB_VneM_XG7 according to claim 1, characterized in that, The bacteriophage vB_VneM_XG7 can maintain relatively stable activity between pH 6 and 11 and between -20°C and 70°C.

4. A microbial preparation, characterized in that, Includes the bacteriophage vB_VneM_XG7 as described in any one of claims 1 to 3.

5. A bactericide, characterized in that, Includes the bacteriophage vB_VneM_XG7 as described in any one of claims 1 to 3.

6. A bacteriophage drug formulation, characterized in that, The active ingredient of the phage drug formulation includes the phage vB_VneM_XG7 as described in any one of claims 1 to 3.

7. The use of the bacteriophage vB_VneM_XG7 according to any one of claims 1 to 3, the microbial preparation according to claim 4, the bactericide according to claim 5, or the bacteriophage drug preparation according to claim 6 in the preparation of products for killing pathogenic Vibrio, characterized in that, The pathogenic Vibrio is Vibrio neocalidonia.

8. The application according to claim 7, characterized in that, The bacteriophage vB_VneM_XG7 lysed 241 PFU / cell of pathogenic Vibrio.

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