Vibrio canbainii bacteriophage VCP13AVCGD9 as well as composition and application thereof

Abstract

The invention discloses a vibrio cantoniensis phage VCP13AVCGD9 as well as a composition and application of the vibrio cantoniensis phage VCP13AVCGD9, the vibrio cantoniensis phage VCP13AVCGD9 is preserved in Guangdong Microbiological Culture Collection Center on November 24, 2025, the preservation number of the vibrio cantoniensis phage VCP13AVCGD9 is GDMCC NO. 67344-B1, and the vibrio cantoniensis phage VCP13AVCGD9 is classified and named as Vibrio campbelli phage VCP13AVCGD9. The vibrio canetinii bacteriophage VCP13AVCGD9 provided by the invention has good acid-base stability, can still keep relatively high titer after being treated for 96 hours under the condition that the pH value is 4.0-8.0, is beneficial to continuously keeping bactericidal activity in a culture water body, and acts on vibrio canetinii in animal gastrointestinal tracts in a targeting manner.

Description

Technical Field

[0001] This application relates to the field of bacteriophage technology, and in particular to Vibrio campbellii bacteriophage VCP13AVCGD9, its compositions and applications. Background Technology

[0002] Vibrio campbellii is a Gram-negative vibrio belonging to the genus Vibrio in the family Vibrioceae. This bacterium is not only a common opportunistic pathogen in aquaculture but can also enter the human food chain through contaminated seafood, causing food safety issues. In aquaculture, Vibrio campbellii can infect various economically important aquatic animals such as shrimp, fish, and shellfish, leading to outbreaks of vibriosis. Typical symptoms include skin ulcers, hepatopancreatic necrosis, enteritis, and systemic septicemia, which can cause large-scale mortality and significant economic losses to the aquaculture industry. Furthermore, Vibrio campbellii can also infect humans, causing gastroenteritis, wound infections, and even septicemia, posing a potential threat to public health.

[0003] Currently, the prevention and control of Vibrio campestris infection still mainly relies on chemical antibiotics (such as fluoroquinolones and tetracyclines) and disinfectants. However, the long-term and widespread use of antibiotics has led to increasingly prominent drug resistance in Vibrio campestris, with multidrug-resistant strains constantly emerging, significantly reducing the effectiveness of traditional drug treatments. At the same time, antibiotic residues also negatively impact the quality and safety of aquatic products and the ecological environment. Furthermore, existing vaccine development still faces technical bottlenecks such as short periods of immune protection, narrow serotype coverage, and limited applicability, making it difficult to meet the diverse and dynamically changing prevention and control needs in actual aquaculture production.

[0004] Bacteriophages, as a class of viruses capable of specifically infecting and lysing bacteria, have become an important direction for the prevention and control of bacterial diseases due to their outstanding advantages such as high host specificity, no impact on the host's normal flora, environmental friendliness, and low likelihood of inducing bacterial resistance. Utilizing bacteriophages to control Vibrio campestris infection can not only achieve precise targeted sterilization and reduce interference with the aquaculture environment and animal microecology, but also provide a new approach for developing green and sustainable aquatic disease control strategies.

[0005] Although bacteriophages have shown promising potential in controlling Vibrio campbellii, currently reported Vibrio campbellii bacteriophages exhibit insufficient tolerance to environmental physicochemical conditions, particularly poor stability to pH fluctuations. In actual aquaculture water, feed processing, or digestive tract environments, pH values ​​often fluctuate significantly. Many existing bacteriophages are easily and rapidly inactivated under acidic or alkaline conditions, resulting in their inability to maintain activity for long periods. This severely impacts the stability of their effectiveness and the range of applicable scenarios in practical applications. Summary of the Invention

[0006] To improve the acid-base stability of bacteriophages, this application provides a strain of Vibrio campbellii bacteriophage VCP13AVCGD9, its composition, and its application.

[0007] In the first aspect, this application provides a strain of Vibrio campeosaphire phage VCP13AVCGD9, which adopts the following technical solution: A strain of Vibrio campbellii bacteriophage VCP13AVCGD9, deposited on November 24, 2025 at the Guangdong Provincial Microbial Culture Collection Center, with accession number GDMCC NO. 67344-B1, and classified as follows: Vibrio campbellii phage VCP13AVCGD9.

[0008] By adopting the above technical solution, the Vibrio campestris phage VCP13AVCGD9 provided in this application exhibits good acid-base stability, maintaining a high titer even after 96 hours of treatment under pH conditions of 4.0-8.0. This characteristic enables it to adapt to common pH fluctuations in aquaculture water, continuously maintaining its bactericidal activity; it also helps the phage survive after passing through the acidic gastric environment of animals, thereby targeting Vibrio campestris in the intestines and improving its environmental adaptability and reliability in practical applications.

[0009] Furthermore, the complete genome sequence of Vibrio campbellii phage VCP13AVCGD9 is shown in SEQ ID NO.1-5.

[0010] Furthermore, the Vibrio campbellii bacteriophage VCP13AVCGD9 has a polyhedral symmetry head and a relatively long tail. The diameter of the head is 56-65 nm, the length of the tail is 107-117 nm, and the diameter of the tail is 6-10 nm.

[0011] Furthermore, the Vibrio campbellii phage VCP13AVCGD9 does not contain virulence genes or undesirable genes.

[0012] Furthermore, the titer of the Vibrio campbellii bacteriophage VCP13AVCGD9 did not change significantly after treatment with chloroform and ether for 2 hours.

[0013] Furthermore, the Vibrio campestris phage VCP13AVCGD9 exhibits relatively stable activity at 55°C and retains 10% activity even after a 24-hour water bath at 75°C. 5 PFU / mL potency.

[0014] Furthermore, the Vibrio campbellii phage VCP13AVCGD9 still exhibits 10% glutathione activity after treatment at pH 13.0 for 8 hours. 3PFU / mL potency.

[0015] Optionally, the Vibrio campei bacteriophage VCP13AVCGD9 can also recognize Vibrio alginolyticus, Vibrio vulnificus, Vibrio flocculus, Vibrio fluvialis, or Vibrio breganum.

[0016] Secondly, this application provides the use of Vibrio campbellii bacteriophage VCP13AVCGD9 in inhibiting Vibrio campbellii.

[0017] Thirdly, this application provides a composition for inhibiting Vibrio campbellii, comprising Vibrio campbellii bacteriophage VCP13AVCGD9.

[0018] Optionally, it also includes one or more of Vibrio campbellii phage VCP11AVCGD4, Citrobacter freundii phage CFP7ACFGC5, and Edwardsiella edodes phage EDP3AEDQD6, wherein the preservation number of Vibrio campbellii phage VCP11AVCGD4 is GDMCC NO. 67343-B1, the preservation number of Citrobacter freundii phage CFP7ACFGC5 is CCTCC NO. M20251640, and the preservation number of Edwardsiella edodes phage EDP3AEDQD6 is GDMCC NO. 66852-B1.

[0019] By adopting the above technical solution, this application provides a composition for inhibiting Vibrio campbellii. This composition uses Vibrio campbellii bacteriophage VCP13AVCGD9 as its core and can selectively combine one or more bacteriophages. It can exert synergistic or synergistic lytic effects against different strains or physiological states of Vibrio campbellii, reducing the risk of control failure due to bacterial bacteriophage resistance mutations, thereby improving the overall strength and reliability of the bactericidal effect. Furthermore, the Citrobacter freundii bacteriophage and Edwardsiella tarda bacteriophage introduced into the composition can target common aquatic pathogens, enabling the composition to not only primarily control Vibrio campbellii but also possess potential inhibitory capabilities against other important pathogens, thus broadening its application scenarios in the control of complex diseases in aquaculture.

[0020] Fourthly, this application provides a feed or feed additive comprising Vibrio campbellii bacteriophage VCP13AVCGD9 or a composition that inhibits Vibrio campbellii.

[0021] Fifthly, this application provides a medicament comprising Vibrio campestris phage VCP13AVCGD9 or a composition that inhibits Vibrio campestris.

[0022] Optionally, the drug includes a pharmaceutically acceptable carrier, and the use of the drug includes application to the surface, mouth, rectum, or pleura of the host being treated, in the form of carrier-borne, concentrated injection, or drug immersion.

[0023] By adopting the above technical solution, this application provides a drug containing Vibrio campbellii bacteriophage VCP13AVCGD9 or a combination thereof, which can be administered in various forms such as carrier delivery, concentrated injection or drug soaking, and can act on different sites such as "body surface, mouth, rectum or pleura", covering common and efficient intervention methods in aquaculture and animal medicine, greatly enhancing the application flexibility and scenario adaptability of the drug.

[0024] In a sixth aspect, this application provides a biological bactericide comprising Vibrio campestris phage VCP13AVCGD9 or a composition that inhibits Vibrio campestris.

[0025] Seventhly, this application provides a kit comprising Vibrio campestris phage VCP13AVCGD9.

[0026] Eighthly, this application provides a method for inhibiting Vibrio campbellii for non-disease diagnosis and treatment purposes, which utilizes the lytic effect of Vibrio campbellii bacteriophage VCP13AVCGD9 or a combination thereof on Vibrio campbellii to inhibit Vibrio campbellii.

[0027] In summary, this application includes at least one of the following beneficial technical effects: 1. The Vibrio campestris phage VCP13AVCGD9 provided in this application exhibits good acid-base stability, maintaining a high titer even after 96 hours of treatment under pH conditions of 4.0-8.0. This characteristic allows it to adapt to common pH fluctuations in aquaculture water, continuously maintaining its bactericidal activity; it also helps the phage survive after passing through the acidic gastric environment of animals, thereby targeting Vibrio campestris in the intestines and improving its environmental adaptability and reliability in practical applications. 2. This application provides a composition for inhibiting Vibrio campbellii. The composition uses Vibrio campbellii bacteriophage VCP13AVCGD9 as its core and can selectively combine one or more bacteriophages. It can exert synergistic or additive lytic effects against different strains or physiological states of Vibrio campbellii, reducing the risk of control failure due to bacterial bacteriophage resistance mutations, thereby improving the overall strength and reliability of the bactericidal effect. Furthermore, the Citrobacter freundii bacteriophage and Edwardsiella tarda bacteriophage introduced into the composition can target common aquatic pathogens, enabling the composition to not only primarily control Vibrio campbellii but also possess potential inhibitory capabilities against other important pathogens, thus broadening its application scenarios in the control of complex diseases in aquaculture. Attached Figure Description

[0028] Figure 1 This is a schematic diagram of a phage plaque from Vibrio campeum in Example 1 of this application; Figure 2 This is a transmission electron microscope image of Vibrio campbellii bacteriophage in Example 1 of this application; wherein, the head diameter of Vibrio campbellii bacteriophage VCP13AVCGD9 in the image is 60 nm, the length of the tail is 112 nm, and the diameter of the tail is 6 nm. Figure 3 This is a distribution diagram of the unique endonuclease cleavage sites of Vibrio campestris phage in Example 2 of this application; Figure 4 This is a phylogenetic tree analysis diagram of the Vibrio campei phage genome from Example 2 of this application; wherein, Vibrio The NCBI Accession number for phage 1.188.A.10N.286.51.A6 is NC_048122; Vibrio The NCBI Accession number for phage 1.188.B.10N.286.51.A6 is MG592555; Vibrio The NCBI Accession number for phage1.169.O.10N.261.52.B1 is NC_048121; Vibrio The NCBI Accession number for phage1.261.O.10N.286.51.A7 is NC_048125; Vibrio The NCBI Accession number for phage1.224.A.10N.261.48.B1 is NC_048124; Figure 5 This is a stability graph of Vibrio campbellii bacteriophage in Example 7 of this application at different temperatures; Figure 6 This is a stability diagram of Vibrio campbellii bacteriophage in Example 7 of this application under different pH conditions; Figure 7 This is a stability graph of Vibrio campbellii bacteriophage in Example 8 of this application before and after preparation under different powder preparation conditions; Figure 8 This is a diagram showing the lytic activity of Vibrio campbellii bacteriophage against Vibrio campbellii in Example 9 of this application. Figure 9This is a diagram illustrating the lytic activity of Vibrio campeosaphireum phage against non-host pathogenic bacteria in Example 10 of this application. In the diagram, A1 represents Vibrio parahaemolyticus, A2 represents Vibrio alginolyticus, A3 represents Vibrio harveyi, A4 represents Vibrio vulnificus, A5 represents Vibrio flocculation, A6 represents Vibrio fluvialis, A7 represents Vibrio splenium, A8 represents Vibrio cholerae, A9 represents Vibrio breganum, A10 represents Citrobacter freundii, A11 represents Edwardsiella tarda, and A12 represents Aeromonas hydrophila. Figure 10 This is a diagram illustrating the lytic activity of Vibrio campestris phage against non-pathogenic beneficial bacteria in Example 11 of this application. In this diagram, B1 represents non-pathogenic rhizobia, B2 represents non-pathogenic Bacillus licheniformis, B3 represents non-pathogenic Bacillus subtilis, B4 represents non-pathogenic Bacillus megaterium, B5 represents Bacillus coagulans, and B6 represents Clostridium butyricum. Figure 11 This is a diagram illustrating the protective effect of Vibrio campestris bacteriophage and its composition on shrimp in Example 14 of this application. Detailed Implementation

[0029] The technical solutions of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0030] The raw materials used in the embodiments of this application can all be obtained from commercially available or preserved institutions, wherein: Vibrio campbellii phage VCP13AVCGD9: Preservation name: Vibrio campbellii phage VCP13AVCGD9 ( Vibrio campbellii Phage VCP13AVCGD9). Collection number: GDMCC NO. 67344-B1. Depository institution: Guangdong Provincial Center for Microbial Culture Collection. Abbreviation: GDMCC. Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Deposit date: November 24, 2025. Vibrio campbellii phage VCP11AVCGD4: Preservation name: Vibrio campbellii phage VCP11AVCGD4 ( Vibrio campbellii Phage VCP11AVCGD4). Collection number: GDMCC NO. 67343-B1. Depository institution: Guangdong Provincial Center for Microbial Culture Collection. Abbreviation: GDMCC. Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Deposit date: November 24, 2025. Citrobacter freundii phage CFP7ACFGC5: Preservation name: Citrobacter freundii phage CFP7ACFGC5 ( Citrobacter freundii Phage CFP7ACFGC5). Collection number: CCTCCNO. M20251640. Depository institution: China Center for Type Culture Collection. Abbreviation: CCTCC. Address: School of Life Sciences, Wuhan University. Deposit date: July 21, 2025. Edwardsiella tarda phage EDP3AEDQD6: Preservation name: Edwardsiella tarda phage EDP3AEDQD6 ( Edwardiella phage EDP3AEDQD6). Collection Center Registration Number: GDMCC NO. 66852-B1. Depository Institution: Guangdong Provincial Center for Microbial Culture Collection. Abbreviation: GDMCC. Address: 5th Floor, Building 59, No. 100 Xianlie Middle Road, Guangzhou. Deposit Date: August 18, 2025; TCBS (thiosulfate citrate bile salt sucrose) agar medium, Haibo Biotechnology; Phage DNA Extraction Kit, Norgen Biotek.

[0031] Preparation Example 1 Preparation of tryptic soy peptone (TSB) liquid culture medium: Mix 15g tryptic peptone, 5g soy peptone, 5g sodium chloride and 1000mL distilled water, and autoclave at 121℃ for 20min to obtain TSB liquid culture medium.

[0032] Preparation Example 2 Preparation of tryptic soy peptone agar (TSA) solid medium: Mix 15g tryptic peptone, 5g soy peptone, 5g sodium chloride, 15g agar and 1000mL distilled water, and autoclave at 121℃ for 20min to obtain TSA solid medium.

[0033] Preparation Example 3 Preparation of TSB semi-solid agar medium: Mix 15g tryptone, 5g soybean peptone, 5g sodium chloride, 7g agar and 1000mL distilled water, and autoclave at 121℃ for 20min to obtain TSB semi-solid agar medium.

[0034] Preparation Example 4 Preparation of Luria-Bertany (LB) liquid medium: Mix 10g of tryptone, 5g of yeast extract, 10g of sodium chloride and 1000mL of distilled water, adjust the pH to 7.0, and autoclave at 121℃ for 20min to obtain LB liquid medium.

[0035] Preparation Example 5 Preparation of LB solid medium: Mix 10g tryptone, 5g yeast extract, 10g sodium chloride, 15g agar and 1000mL distilled water, adjust the pH to 7.0, and autoclave at 121℃ for 20min to obtain LB solid medium.

[0036] Preparation Example 6 Preparation of LB semi-solid agar medium: Mix 10g tryptone, 5g yeast extract, 10g sodium chloride, 7g agar and 1000mL distilled water, adjust the pH to 7.0, and autoclave at 121℃ for 20min to obtain semi-solid agar medium.

[0037] Preparation Example 7 Preparation of SM buffer: Mix 5.8g sodium chloride, 2g magnesium sulfate, 50mL 1mol / L tris(hydroxymethyl)aminomethane hydrochloride, 0.25g gelatin and 1000mL distilled water, and autoclave at 121℃ for 20min to obtain SM buffer.

[0038] Preparation Example 8 Preparation of logarithmic phase Vibrio campbellii AVCGD9 bacterial suspension: Urban sewage collected from Guangzhou, Guangdong Province, was serially diluted with sterile physiological saline. Each dilution was then spread onto TCBS agar and incubated at 30°C for 10 h. Typical single colonies were picked and inoculated onto TCBS agar and incubated at 30°C for 10 h. This process was repeated at least three times until morphologically uniform single colonies were obtained. Single colonies were selected for 16S rRNA gene amplification and sequencing. The sequencing results were compared with BLAST to confirm the presence of Vibrio campbellii, and the strain was named Vibrio campbellii AVCGD9. A single Vibrio campbellii AVCGD9 colony was picked and inoculated into 3 mL of TSB liquid medium obtained in Example 1, and incubated at 30°C for 6 h to obtain the logarithmic phase Vibrio campbellii AVCGD9 bacterial suspension.

[0039] Preparation Example 9 Preparation of host bacterial suspension: A single colony of Vibrio campbellii AVCGD9 obtained in Preparation Example 8 was picked and inoculated into a test tube containing 3 mL of LB liquid medium obtained in Preparation Example 4. The culture was carried out at 30°C and 150 rpm for 8 h with shaking to obtain the host bacterial suspension.

[0040] Example 1: Screening, Separation and Purification Samples were collected from urban sewage in Guangzhou, Guangdong Province. The collected samples were centrifuged at 5000 rpm for 10 min and filtered through a 0.22 μm filter membrane to obtain a collection solution. 10 mL of the collection solution was mixed with the LB liquid medium obtained in Preparation Example 4 at a 1:1 ratio. 1 mL of *Vibrio campestris* AVCGD9 logarithmic phase bacterial suspension obtained in Preparation Example 8 was added, and the mixture was incubated overnight at 30°C. After centrifugation at 5000 rpm for 10 min, the supernatant was filtered through a 0.22 μm filter membrane to obtain the enriched solution. 0.5 mL of *Vibrio campestris* AVCGD9 logarithmic phase bacterial suspension obtained in Preparation Example 8 was added to 5 mL of LB semi-solid agar medium obtained in Preparation Example 6 at 48°C and mixed thoroughly. The mixture was then poured onto the LB solid medium obtained in Preparation Example 5 to obtain a double-layer plate. 10 μL of the enriched solution was dropped onto the solidified double-layer plate, air-dried under aseptic conditions, and incubated overnight at 30°C to obtain bacteriophage spotting.

[0041] Pick a phage spot and place it in 1 mL of the SM buffer obtained in Preparation Example 7. Shake at 150 rpm for 15 min. Then, perform serial dilutions with SM buffer to obtain 1×10⁻⁶ phages. -2 1×10 -4 1×10 -6 1×10 -8 For the dilution buffer, add 0.5 mL of each serially diluted Vibrio campestris AVCGD9 logarithmic phase culture obtained in Preparation Example 8, mix thoroughly, then add to 5 mL of LB semi-solid medium obtained in Preparation Example 6 and mix again. Pour onto LB solid medium obtained in Preparation Example 5, and after the semi-solid medium solidifies, incubate overnight at 30°C. Repeat this step 3-5 times to finally obtain single phage plaques of uniform morphology and size on double-layer plates (see...). Figure 1 This refers to the purification of the bacteriophage, which is named bacteriophage VCP13AVCGD9.

[0042] A single plaque of purified bacteriophage was picked and placed in a logarithmic phase culture of Vibrio campestris AVCGD9 obtained in Preparation Example 8, and added to 50 mL of LB liquid medium obtained in Preparation Example 4. The culture was incubated at 30°C for 8 h, then centrifuged at 8000 rpm for 10 min, and the supernatant was filtered through a 0.22 μm filter membrane to obtain a pure culture of bacteriophage VCP13AVCGD9.

[0043] Depend on Figure 1 It can be seen that the purified bacteriophages are uniform in morphology and size, with clear and regular edges, indicating that the single bacteriophage VCP13AVCGD9 virus strain has been successfully purified.

[0044] Pure culture medium of bacteriophage VCP13AVCGD9 was irradiated with a transmission electron microscope, and the results were observed. Figure 2 .Depend on Figure 2It is known that bacteriophage VCP13AVCGD9 has a polyhedral symmetrical head and a relatively long tail. The head has a diameter of 56-65 nm, the tail has a length of 107-117 nm, and the tail has a diameter of 6-10 nm.

[0045] Example 2: Genome extraction, sequencing, and whole-genome analysis Nucleic acid from bacteriophage VCP13AVCGD9 was extracted using a bacteriophage DNA extraction kit and sequenced. The sequencing results showed that the genome of bacteriophage VCP13AVCGD9 is a linear double-stranded DNA, with nucleotide sequences of SEQ ID NO.1-5, a total genome length of 72565 bp, and a G+C content of 44.3%.

[0046] Genome-wide prediction of bacteriophage VCP13AVCGD9 using PHASTEST (https: / / phastest.ca / ) revealed 110 bacteriophage gene sequences and 5 bacterial genome sequences. For structural details, see [link to PHASTEST]. Figure 3 .

[0047] Phylogenetic analysis of the VCP13AVCGD9 genome was performed in NCBI, and the results are shown below. Figure 4 .Depend on Figure 4 It can be seen that the bacteriophage VCP13AVCGD9 is genetically closest to the bacteriophage VCP13AVCGD9. Vibrio phage1.188.A.10N.286.51.A6 (NC_048122) and Vibrio Phage 1.188.B.10N.286.51.A6(MG592555).

[0048] Combining the unique genomic information of bacteriophage VCP13AVCGD9 with its host characteristics and morphological features under transmission electron microscopy, and identifying it according to the classification criteria established by the International Committee on Virus Taxonomy, the results show that bacteriophage VCP13AVCGD9 belongs to the class Caudoviricetes, family Schitoviridae, and genus Mukerjeevirus. Given that the host bacteria from which bacteriophage VCP13AVCGD9 was isolated is... Vibrio campbellii Therefore, bacteriophage VCP13AVCGD9 is Vibrio campbellii Bacteriophage.

[0049] Example 3: Detection of virulence gene or defective gene deletion The 103 virulence genes described in patent CN120464583A were tested. The results showed that Vibrio campbellii phage VCP13AVCGD9 does not contain virulence genes or harmful genes, and therefore cannot encode proteins that may cause potential health risks. Therefore, Vibrio campbellii phage VCP13AVCGD9 will not affect the health of humans or animals.

[0050] Example 4: Determination of Optimal Multiplicity of Infection The pure culture medium of phage VCP13AVCGD9 obtained in Example 1 was serially diluted 10-fold using SM buffer to obtain diluted phage VCP13AVCGD9. The 10⁻¹⁰ m² plate assay was performed. -5 up to l0 -8 The phage titer is calculated based on the dilution. Each dilution requires three replicates. After incubation, plaques are counted and the titer is calculated. Phage titer (PFU / mL) = average number of plaques × dilution factor.

[0051] The host bacterial suspension obtained in Preparation Example 9 was transferred to 10 mL of LB liquid medium obtained in Preparation Example 4 at a ratio of 1:100. The culture was incubated at 30°C and 150 rpm with shaking until the early logarithmic growth phase. Then, pure culture medium of phage VCP13AVCGD9 obtained in Example 1 was added at ratios of 1000:1, 100:1, 10:1, 1:1, 1:10, 1:100, and 1:1000, respectively. LB liquid medium was added to ensure the total volume of each tube was the same. The tubes were incubated at 30°C and 150 rpm for 8 hours. After incubation, the tubes were centrifuged at 5000g for 10 min, and the supernatant was collected to determine the phage titer. Double-sample culture was performed at each point, and the average value was taken. The experiment was repeated three times. The results are shown in Table 1.

[0052] Table 1. Titer of Vibrio campbellii bacteriophage VCP13AVCGD9 at different multiplicity of infection

[0053] As shown in Table 1, the highest titer of Vibrio campei phage VCP13AVCGD9 is 4 × 10⁻⁶. 10 At a PFU / mL level, the multiplicity of infection was 0.001, indicating that Vibrio campestris phage VCP13AVCGD9 has a strong infectivity.

[0054] Example 5: Toxicological Experiment Eighty healthy, specific pathogen-free zebrafish, approximately 3-4 cm in length, were selected and acclimatized for 3 days. They were then randomly divided into two groups (phage group and control group), with 40 fish in each group and a stocking density of 20 fish / 50L. The phage VCP13AVCGD9 dilution obtained in Example 4 was diluted using the SM buffer solution obtained in Preparation Example 7, yielding a titer of 1×10⁻⁶. 8Phage dilution A (PFU / mL) was added to the phage group. 100 μL of phage dilution A was added to the control group, and an equal volume of sterile saline was added to the control group. Each group was fed twice daily, with a 20% water change per day. After each water change, phage dilution A and sterile saline were added proportionally to maintain the final concentration. The zebrafish were fed continuously for 15 days, and their condition was observed and recorded.

[0055] One hundred and eighty healthy, specific pathogen-free Litopenaeus vannamei shrimp, each weighing approximately 1g, were selected and acclimatized for 3 days. They were then randomly divided into two groups (phage group and control group), with 90 shrimp in each group and a stocking density of 30 shrimp / 50L. The phage group was given 100μL of phage dilution A, while the control group was given an equal volume of sterile saline. Each group was fed twice daily, with a 20% water change per day. After each water change, the phage titer was replenished at a rate of 1×10⁻⁶. 8 The phage dilution at PFU / mL and sterile saline were used to maintain the final concentration. The shrimp were fed continuously for 15 days, and the condition of Litopenaeus vannamei was observed and recorded.

[0056] Twenty healthy, age- and weight-less pathogen-free experimental mice (half male and half female) were selected and, after acclimatization for 3 days, were randomly divided into two groups (phage group and control group), with 10 mice in each group (5 males and 5 females). The phage VCP13AVCGD9 dilution obtained in Example 4 was diluted with the SM buffer obtained in Preparation Example 7 to obtain a titer of 1×10⁻⁶. 10 Phage dilution B (PFU / mL) was administered. The phage group received 100 μL of phage dilution B via tail vein injection, while the control group received an equal volume of sterile saline via tail vein injection. Food intake, activity levels, body temperature, weight changes, and survival were observed. Ten days later, the mice were euthanized by cervical dislocation, and their internal organs were examined.

[0057] Forty healthy, age- and weight-similar, specific pathogen-free experimental mice (half male and half female) were selected and, after three days of acclimatization, were randomly divided into four groups (phage group 1, phage group 2, phage group 3, and control group), with 10 mice in each group (5 males and 5 females). The phage VCP13AVCGD9 dilution obtained in Example 4 was diluted using the SM buffer obtained in Preparation Example 7 to obtain a titer of 2 × 10⁻⁶. 9 Phage dilution C at a titer of PFU / mL was administered orally to phage groups 1, 2, and 3 at titers of 0.2 mL / mouse, 0.5 mL / mouse, and 1 mL / mouse, respectively. The control group received an equal volume of physiological saline. The mice were observed for 15 consecutive days. The mice were then euthanized by cervical dislocation, and their internal organs were examined.

[0058] The results showed that, in all the above treatment groups, immersion in high concentrations of Vibrio campestris phage VCP13AVCGD9 had no effect on the survival of zebrafish and Litopenaeus vannamei, with no morbidity or mortality. Intravenous injection of a large dose of Vibrio campestris phage VCP13AVCGD9 into the tail vein had no effect on the daily behavior of mice; during the experiment, mice did not exhibit any clinical symptoms such as lethargy, rough coat, decreased appetite, weight loss, or death, and no abnormalities were found in the tissues and internal organs upon anatomical examination. Oral administration of different doses of Vibrio campestris phage had no effect on the daily behavior of mice, with no symptoms such as depression, fever, vomiting, or diarrhea. No abnormalities were found in the internal organs upon anatomical examination of mice in any treatment. This indicates that the Vibrio campestris phage VCP13AVCGD9 of this application is biosafety and can be used as a feed additive or pharmaceutical.

[0059] Example 6: Chloroform and Ether Tolerance Chloroform sensitivity test: The potency obtained in Example 4 was 5.5 × 10⁻⁶. 8 950 μL of PFU / mL phage dilution was mixed with 50 μL of chloroform and placed in a sterile EP tube as the test group. 50 μL of SM buffer was placed in a sterile EP tube, and 950 μL of the titer obtained in Example 4 (5.5 × 10⁻⁶) was added to it. 8 A phage dilution of PFU / mL was used as a control group. After the above treatment was thoroughly mixed, the mixture was incubated at 4°C overnight, and the titer was determined. The results are shown in Table 2.

[0060] Ether sensitivity test: The titer obtained in Example 4 was 5.5 × 10⁻⁶. 8 800 μL of PFU / mL phage dilution was mixed with 200 μL of diethyl ether and placed in a sterile EP tube as the test group; 200 μL of SM buffer was placed in a sterile EP tube, and 800 μL of the titer obtained in Example 4 (5.5 × 10⁻⁶) was added to it. 8 The phage dilution of PFU / mL was used as the control group. After the above treatment was thoroughly mixed, the mixture was shaken in an ice bath for 2 hours, centrifuged at 3000 r / min for 15 min, and the titer was determined by taking the aqueous phase. The results are shown in Table 2.

[0061] Table 2. Effects of chloroform and ether treatments on the titer of bacteriophage VCP13AVCGD9

[0062] Table 2 shows that after treatment with chloroform and ether for 2 hours, the titer of Vibrio campestris phage VCP13AVCGD9 did not change significantly compared with the control group, indicating that phage VCP13AVCGD9 has good tolerance to chloroform and ether. Chloroform is lipid-soluble, which indirectly suggests that the capsid of phage VCP13AVCGD9 does not contain lipids.

[0063] Example 7: Physicochemical Stability Thermal stability: 30 mL of the pure culture medium of phage VCP13AVCGD9 obtained in Example 4 was dispensed into sterile EP tubes, divided into three equal portions of 10 mL each, and incubated in water baths at 4℃, 55℃, 65℃, and 75℃ for 2 h, 24 h, and 48 h, respectively. A control group was also included, receiving an equal volume of pure phage VCP13AVCGD9 culture medium and incubating in a water bath at 4℃ for 2 h, 24 h, and 48 h. After the incubation period, the sample tubes were removed and immediately placed in an ice bath to cool. After appropriate dilution, the phage titer was determined using the double-layer plate method. The experiment was repeated three times, and the results are shown below. Figure 5 .

[0064] Depend on Figure 5 It can be seen that the activity of Vibrio campbellii phage VCP13AVCGD9 in the experimental group was relatively stable at 55℃. Vibrio campbellii phage VCP13AVCGD9 exhibits good heat resistance; compared with the control, its titer decreased by only two orders of magnitude after treatment at 65℃ for 48 hours, and it still maintained 10% titer after a 24-hour water bath at 75℃. 5 PFU / mL potency.

[0065] Acid-base stability: The potency obtained in Example 4 was 5.5 × 10⁻⁶. 8 Phage dilutions (PFU / mL) were adjusted to pH values ​​between 1.0 and 14.0. After treatment for 1 h, 4 h, 8 h, 24 h, and 96 h, the titers were measured. Results are shown below. Figure 6 .

[0066] Depend on Figure 6 As shown, Vibrio campbellii phage VCP13AVCGD9 still retained 10% saturation after treatment at pH 13.0 for 8 hours. 3 The PFU / mL titer remained stable and high after treatment at pH 4.0-8.0 for 96 h and at pH 3.0-10.0 for 24 h. Vibrio campestris phage VCP13AVCGD9 exhibited good acid and alkali tolerance, especially alkali tolerance.

[0067] Example 8: Bioactivity Test of Spray Powder and Lyophilized Powder Bioactivity test of the spray powder: The potency obtained by dissolving 2% starch (w / v) and 4% sorbitol (w / v) in Example 4 was 5.5 × 10⁻⁶. 8Phage was coated by incubating a PFU / mL phage dilution at 30°C and 180 rpm for 2 hours. The coated phage solution was then sprayed onto defatted rice bran carrier at a 1:4 (w / w) ratio, stirred to ensure homogeneity, dried at 50°C for 2 hours, and passed through a 40-mesh sieve. After preparation, samples were taken and the phage titer was determined using the double-layer plate method. The experiment was repeated three times.

[0068] Bioactivity test of the lyophilized powder: 2% starch (w / v) and 4% sorbitol (w / v) were dissolved in a solution with a potency of 5.5 × 10⁻⁶ obtained in Example 4. 8 Phage was coated by incubating the phage solution in a PFU / mL phage dilution at 30℃ and 180 rpm for 2 h. The coated phage solution was then added to a material tray, freeze-dried under vacuum for 50 h, and finally pulverized through a 40-mesh sieve. After preparation, samples were taken and the phage titer was determined using the double-layer plate method. The experiment was repeated three times, and the results are shown below. Figure 7 .

[0069] Depend on Figure 7 It can be seen that there is no significant difference in titer between the preparation of Vibrio campbellii phage VCP13AVCGD9 into spray powder and lyophilized powder, respectively, indicating that the phage has good tolerance and stability to the preparation conditions of spray powder and lyophilized powder.

[0070] Example 9: Host Bacterial Lysis Experiment Single colonies of Vibrio campeosa isolated from shrimp, fish, and shellfish were selected, totaling 126 strains. Each strain was inoculated into test tubes containing 3 mL of LB liquid medium obtained in Preparation Example 4 and cultured at 30°C for 4 hours to obtain bacterial suspensions. The phage VCP13AVCGD9 dilution obtained in Example 4 was diluted using the SM buffer obtained in Preparation Example 7 to obtain a titer of 1×10⁻⁶. 7 Phage dilution D (PFU / mL). 300 μL of bacterial culture from each bacterium was mixed with the LB semi-solid medium obtained in Example 6 and spread onto ordinary agar plates. 5 μL of phage dilution E was then added to each plate. After air drying, the plates were incubated at 30°C for 8 hours. The results are shown in Table 3. Figure 8 .

[0071] Table 3. Lytic activity of Vibrio campbellii bacteriophage VCP13AVCGD9 against Vibrio campbellii isolates from different animals.

[0072] From Table 3 and Figure 8It is known that the Vibrio campei bacteriophage VCP13AVCGD9 of this application has a wide host range, can recognize Vibrio campei isolated from different animal sources, and has a lysis rate of over 96%, making it well applicable in the preparation of biocides for lysing Vibrio campei.

[0073] Example 10: Lysis test of non-host pathogenic bacteria 228 single colonies of non-host pathogenic bacteria were selected, including Vibrio parahaemolyticus (V. parahaemolyticus). Vibrio parahemolyticus ) 20 strains, Vibrio alginolyticus ( Vibrio alginolyticus ) 20 strains, Vibrio harveyi ( Vibrio harveyi 20 strains, Vibrio vulnificus ( Vibrio vulnificus ) 20 strains, Turbovibrio ( Vibrio scophthalmi ) 20 strains, Vibrio fluvialis ( Vibrio fluvialis ) 20 strains, Vibrio splendidus ( Vibrio splendidus ) 20 strains, Vibrio cholerae ( Vibrio cholerae ) 20 strains, Vibrio bregenii ( Vibrio breoganii ) 8 strains, Citrobacter freundii ( Citrobacter freundii 20 strains, Edwardsiella tarda ( Edwardiella 20 strains and Aeromonas hydrophila ( Aeromonas hydrophila 20 strains were prepared. The tested bacterial strains were inoculated into test tubes containing 3 mL of the TSB liquid medium obtained in Preparation Example 1, and cultured under optimal conditions until the logarithmic growth phase. 300 μL of each bacterial culture was mixed with LB semi-solid medium obtained in Preparation Example 6 and plated onto ordinary agar plates. 5 μL of the culture with a titer of 1 × 10⁻⁶ obtained in Example 9 was also mixed. 7 PFU / mL phage dilution D was dropped onto each plate. After air drying, the plates were incubated overnight under the optimal culture conditions for each tested bacterium. The results are shown in Table 4. Figure 9 .

[0074] Table 4. Lytic ability of Vibrio campbellii bacteriophage VCP13AVCGD9 against non-host pathogenic bacteria.

[0075] From Table 4 and Figure 9 It is known that the Vibrio campei phage VCP13AVCGD9 of this application can recognize some Vibrio alginolyticus, Vibrio vulnificus, Vibrio flocculationus, Vibrio fluvialis, and Vibrio breganum, indicating that the tested phage has a wide range of intrageneric lytic capabilities.

[0076] Example 11: Lysis test of non-pathogenic beneficial bacteria Sixty-four single colonies of non-pathogenic beneficial bacteria were selected, including 12 strains of non-pathogenic rhizobium, 11 strains of non-pathogenic Bacillus licheniformis, 9 strains of non-pathogenic Bacillus subtilis, 12 strains of non-pathogenic Bacillus megaterium, 12 strains of Bacillus coagulans, and 8 strains of Clostridium butyricum. The above-mentioned test strains were inoculated into test tubes containing 3 mL of TSB liquid medium obtained in Preparation Example 1, and cultured under optimal conditions to the logarithmic growth phase to obtain bacterial suspensions for each strain. 300 μL of the bacterial suspension was mixed with LB semi-solid medium obtained in Preparation Example 6 and plated onto ordinary agar plates. 5 μL of the 1×10⁻⁶ titer obtained in Example 9 was also mixed. 7 PFU / mL phage dilution D was dropped onto each plate. The plates were then incubated overnight under optimal conditions for each tested bacterium. Results are shown in the table below. Figure 10 .

[0077] Depend on Figure 10 It is known that the Vibrio campei phage VCP13AVCGD9 of this application could not recognize any of the 64 tested non-pathogenic beneficial bacteria, indicating that the tested phage had no damaging effect on the beneficial bacterial microbial community.

[0078] Example 12: Preparation of the composition Vibrio campei phage VCP11AVCGD4, Citrobacter freundii phage CFP7ACFGC5, and Edwardsiella tarda phage EDP3AEDQD6 were cultured and activated, and then diluted with SM buffer to obtain titers of 2×10⁻⁶ for all phages. 9 A dilution of PFU / mL.

[0079] The valence obtained in Example 5 was 2 × 10. 9 Equal volumes of PFU / mL phage dilution C and Vibrio campbellii phage VCP11AVCGD4 dilution were mixed to obtain composition 1.

[0080] Equal volumes of phage dilution C and Citrobacter freundii phage CFP7ACFGC5 dilution were mixed uniformly to obtain composition 2.

[0081] Equal volumes of phage dilution C and Edwardsiella edodes phage EDP3AEDQD6 dilution were mixed uniformly to obtain composition 3.

[0082] Equal volumes of phage dilution C, Vibrio campbellii phage VCP11AVCGD4 dilution, Citrobacter freundii phage CFP7ACFGC5 dilution, and Edwardsiella tarda phage EDP3AEDQD6 dilution were mixed to obtain composition 4.

[0083] The phage dilution C and 200 mg / L hydrogen peroxide aqueous solution were mixed in equal volumes at a ratio of 1:1 to obtain composition 5.

[0084] Example 13: Bactericidal effect of Vibrio campestris bacteriophage VCP13AVCGD9 and its composition in liquid. The colony count was performed using a dilution-spreading method. *Vibrio campbellii* was cultured on the TSA solid medium obtained in Preparation Example 2 at 30°C for 24 hours, and the colony count was recorded. The logarithmic phase AVCGD9 culture of *Vibrio campbellii* obtained in Preparation Example 9 was aliquoted into different test tubes and diluted with an equal volume of TSB liquid medium obtained in Preparation Example 1 to a final concentration of 2.4 × 10⁻⁶. 3 CFU / mL. The above test tubes were inoculated with different concentrations of pure culture medium of phage VCP13AVCGD9, so that the concentration of Vibrio campestris phage VCP13AVCGD9 in the samples was 1×10⁻⁶ CFU / mL. 2 PFU / mL, 1×10 3 PFU / mL, 1×10 4 PFU / mL, 1×10 5 PFU / mL, 1×10 6 The concentration of PFU / mL was determined and the samples were divided into five groups: Experiment 1, Experiment 2, Experiment 3, Experiment 4, and Experiment 5. A control group and a blank control group were also included. The control group received an equal volume of PFU / mL at a final concentration of 2.4 × 10⁻⁶. 3 The control group was given an equal volume of physiological saline, with CFU / mL of Vibrio campestris. The residual amount of Vibrio campestris was measured at 2 h and 4 h, and the results are shown in Table 5.

[0085] Table 5. Bactericidal effect of different concentrations of Vibrio campestris phage VCP13AVCGD9 in liquids.

[0086] Table 5 shows that the concentration of Vibrio campbellii bacteriophage VCP13AVCGD9 is 10. 5 -10 6 At a concentration of PFU / mL, Vibrio campbellii phage VCP13AVCGD9 showed the best bactericidal effect against Vibrio campbellii in TSB liquid medium, with a kill rate of over 93%, indicating that Vibrio campbellii phage VCP13AVCGD9 has the potential to be used as a biocide.

[0087] The final concentrations of Vibrio campbellii phage VCP13AVCGD9 in compositions 1, 3, 4, and 5 obtained in Example 12 were diluted to 1 × 10⁻⁶ using TSB liquid medium. 6 PFU / mL. The final concentration was 2.4 × 10⁻⁶. 3 Vibrio campbellans AVCGD9 at a final concentration of 1×10⁻⁶ CFU / mL was inoculated with the bacteria. 6Compositions 1, 2, 3, 4, and 5 of Vibrio campestris phage VCP13AVCGD9 at PFU / mL concentrations were administered. A control group and a blank group were set up. The control group was given a final concentration of 2.4 × 10⁻⁶ PFU / mL. 3 The control group was given an equal volume of physiological saline at CFU / mL. The residual amount of Vibrio campeum was measured after 2 h and 4 h, and the results are shown in Table 6.

[0088] Table 6. Bactericidal effect of different concentrations of Vibrio campestris phage VCP13AVCGD9 compositions in liquid.

[0089] Table 6 shows that the concentration of Vibrio campbellii bacteriophage VCP13AVCGD9 is 1×10⁻⁶. 6 At PFU / mL, the composition containing it not only exhibits good bactericidal effect but also shows no antagonistic effect on other components. The Vibrio campeosa phage VCP13AVCGD9 composition has the potential to be used as a biocide.

[0090] Example 14: Protective effect of Vibrio campbellii bacteriophage VCP13AVCGD9 and its composition on Litopenaeus vannamei. Specific pathogen-free Litopenaeus vannamei shrimp weighing approximately 5g were selected and acclimatized for 3 days. They were then divided into 6 groups (single phage VCP13AVCGD9 group, composition 1 group, composition 2 group, composition 3 group, composition 4 group, composition 5 group, control group, and blank group), with 50 shrimp in each group. A final concentration of 2.4 × 10⁻⁶ was added to the water in each of the single phage VCP13AVCGD9 group, composition 1 group, composition 2 group, composition 3 group, composition 4 group, composition 5 group, and control group. 5 CFU of Vibrio campeum was added to the single phage VCP13AVCGD9 group after 3 hours, with a final concentration of 1×10⁻⁶ obtained in Example 4. 5 The phage VCP13AVCGD9 dilution was added to Composition 1, with a final concentration of 1×10⁻⁶ PFU / mL. 5 Composition 1 (PFU / mL) and Composition 2 (containing Vibrio campestris phage VCP13AVCGD9 at a final concentration of 1×10⁻⁶) 5 Composition 2 and Composition 3, with PFU / mL, contain Vibrio campestris phage VCP13AVCGD9 at a final concentration of 1×10⁻⁶. 5 Compositions 3 and 4, containing PFU / mL, were supplemented with Vibrio campestris phage VCP13AVCGD9 at a final concentration of 1×10⁻⁶. 5 Composition 4 and Composition 5, with a PFU / mL concentration, contain Vibrio campestris phage VCP13AVCGD9 at a final concentration of 1×10⁻⁶.5 Composition 5 (PFU / mL), Compositions 1, 2, 3, 4, and 5 were prepared using the same methods as in Example 12; the control group was replaced with an equal volume of sterile SM buffer. The blank group water was not treated in any way. Each group was fed twice daily, and on day 5 of the experiment, 20% of the water was changed. After the water change, the corresponding phage or composition filtrate and sterile SM buffer were replenished proportionally to maintain the final concentration. The shrimp were fed continuously for 10 days, and their condition was observed and recorded.

[0091] Depend on Figure 11 It was found that by day 10 of the experiment, 2 Litopenaeus vannamei shrimp in the blank group died; only 12 Litopenaeus vannamei shrimp survived in the control group, with a mortality rate of 76%; the number of surviving Litopenaeus vannamei shrimp in the single phage VCP13AVCGD9 group, composition 1 group, composition 2 group, composition 3 group, composition 4 group, and composition 5 group were all above 41, with a mortality rate of less than 18%. Among them, the highest number of surviving Litopenaeus vannamei shrimp was found in composition 4 group, with a survival rate of 92%. This indicates that the Vibrio campbellii bacteriophage VCP13AVCGD9 and its compositions have a significant protective effect on Litopenaeus vannamei shrimp and can be used as a biological bactericide to effectively kill Vibrio campbellii in aquaculture water and protect aquatic animals, thereby improving their survival rate.

[0092] Example 15: Preparation and Use of the Reagent Kit The phage VCP13AVCGD9 dilution buffer obtained in Example 4 was diluted with the SM buffer obtained in Preparation Example 7 to obtain a titer of 1×10⁻⁶. 9 Phage dilution E at PFU / mL. The kit contains 10 mL of phage dilution with a titer of 1×10⁻⁶. 9 1L of phage dilution E was used to prepare LB liquid culture medium obtained in Example 4, 1L of LB solid culture medium obtained in Example 5, and 1L of LB semi-solid culture medium obtained in Example 6.

[0093] The kit usage instructions are as follows: Take 1×10⁻⁶ units of reagent. 9 PFU / mL phage dilution E was used to determine the lysis profile of the tested phages using the double-layer plate drop method. Single colonies of the test strain were picked and inoculated into LB liquid medium, shaken and cultured for 8 hours to obtain the test strain culture. 300 μL of the test strain culture was mixed with 5 mL of LB semi-solid medium and plated onto LB solid plates. 10 μL of a 1×10⁻⁶ titer was then used. 9 Pipette PFU / mL phage dilution E onto a plate. After air drying, incubate for 8 hours and observe the results to determine whether the target strain is present.

[0094] The above are all preferred embodiments of this application, and are not intended to limit the scope of protection of this application. Therefore, all equivalent changes made in accordance with the structure, shape and principle of this application should be covered within the scope of protection of this application.

Claims

1. A strain of Vibrio camberii phage VCP13AVCGD9, characterized in that, The Vibrio campbellii phage VCP13AVCGD9 is deposited with the Guangdong Microbial Culture Collection Center on November 24, 2025, and has a deposit number of GDMCC NO. 67344-B1 and a classification name of Vibrio campbellii phage VCP13AVCGD9.

2. A composition for inhibiting Vibrio campbellii, characterized in that, The Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1.

3. The composition for inhibiting Vibrio campbellii according to claim 2, wherein Further included are one or more of the Vibrio campbellii phage VCP11AVCGD4, Citrobacter freundii phage CFP7ACFGC5, and Edwardsiella phage EDP3AEDQD6, the Vibrio campbellii phage VCP11AVCGD4 having a deposit number of GDMCC NO. 67343-B1, the Citrobacter freundii phage CFP7ACFGC5 having a deposit number of CCTCC NO. M20251640, and the Edwardsiella phage EDP3AEDQD6 having a deposit number of GDMCC NO. 66852-B1.

4. A feed additive, characterized in that, The Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1 or the composition as claimed in any one of claims 2-3.

5. A medicament, characterized by comprising a compound of the formula (I) or a pharmaceutically acceptable salt thereof. The Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1 or the composition as claimed in any one of claims 2-3.

6. The medicament according to claim 5, characterized in that, The drug includes a pharmaceutically acceptable carrier, and the use of the drug includes applying to the surface of the host to be controlled, the mouth, the rectum, or the inside of the pleura in the form of carrier carrying, concentrated injection, or medicament soaking.

7. A biocide, characterized in that, The Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1 or the composition as claimed in any one of claims 2-3.

8. A kit characterized in that, The Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1 or the composition as claimed in any one of claims 2-3.

9. A method of inhibiting Vibrio campbellii for non-disease diagnostic and therapeutic purposes, characterized by, The lysis of the Vibrio campbellii by the Vibrio campbellii phage VCP13AVCGD9 as claimed in claim 1 or the composition as claimed in any one of claims 3-4 inhibits the Vibrio campbellii.

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