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Bacteriophage and application thereof

A bacteriophage, microbial strain technology, applied in the direction of bacteriophage, virus/phage, medical raw materials derived from virus/phage, etc., can solve the problem of high mortality, reduce infection, prevent further deterioration, and treat fish septicemia.

Pending Publication Date: 2020-01-17
YUNNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Aeromonas hydrophila can also cause diarrhea and sepsis in pigs, chickens, ducks, and rabbits, with high mortality

Method used

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  • Bacteriophage and application thereof
  • Bacteriophage and application thereof
  • Bacteriophage and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Isolation, purification and propagation of phage AhyV-DH1

[0030] The host bacteria used in the present invention is Aeromonas hydrophila 4572 isolated from Dianchi Lake by the inventors.

[0031] On September 5, 2018, a water sample was collected from Dianchi Lake, Kunming City, Yunnan Province, and immediately brought back to the laboratory. Added 100 mL of TSB liquid medium and 150 μL of Aeromonas hydrophila 4572 (OD600=0.8) to 50 mL of water sample, 120 rpm, 30°C shaker culture, after 24h, take 50mL of the above culture solution and centrifuge at 4000rpm for 30min, take the supernatant and filter it through a 0.22μm filter membrane, dilute 1000 times with TSB liquid medium, use the double-layer plate method, take the diluted Mix 100 μl of the solution with 200 μl of the host bacterial suspension grown to the logarithmic phase, absorb at 30°C for 10 minutes, then mix evenly with the TSB medium containing 0.75% agar after cooling to 45°C, and pour it on th...

Embodiment 2

[0034] Example 2: Optimal multiplicity of infection (MOI) of phage AhyV-DH1

[0035] Mix 0.5mL Aeromonas hydrophila 4572 culture solution with 0.5mL bacteriophage AhyV-DH1 suspension, so that the multiplicity of infection is 1000, 100, 10, 1, 0.1, 0.01, 0.001, and the difference is determined by double-layer plate method. The phage titer after 3.5 hours of infection under the multiplicity of infection, the infection ratio with the highest titer is the optimal multiplicity of infection; as shown in Table 1, the optimal multiplicity of infection of phage AhyV-DH1 is 0.01.

[0036] Table 1 Infection effect of phage AhyV-DH1 with different MOI

[0037] .

Embodiment 3

[0038] Embodiment 3: One-step growth curve of phage AhyV-DH1

[0039] The phage and Aeromonas hydrophila 4572 were mixed according to the multiplicity of infection of 0.01, and cultured at 120 rpm at 30°C. After infection, 0min, 10min, 20min, 30min, 40min, 50min, 60min, 80min, 100min, 120min, 140min, Take 200 μl of infection solution at 160 min, 180 min, 200 min, and 220 min respectively, and use the double-layer plate method to measure the phage titer in the infection solution. With the post-infection time as the abscissa and the phage titer as the ordinate, draw a one-step growth curve. Such as image 3 As shown, the incubation period of phage AhyV-DH1 is 50 minutes, and the burst volume is 100 PFU / cell.

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Abstract

The invention discloses bacteriophage AhyV-DH1. The preservation number of the bacteriophage AhyV-DH1 in China General Microbiological Culture Collection Center is CGMCC No. 17581. The bacteriophage AhyV-DH1 has a splitting effect on aeromonas hydrophila from different sources. When the bacteriophage AhyV-DH1 is used to control aeromonas hydrophila infecting fish, experiment results show that thebacteriophage AhyV-DH1 can evidently lower the number aeromonas hydrophila at both fish body level and cell level and increase fish survival rate. The bacteriophage AhyV-DH1 can reduce the occurrenceprobability of antibiotic-resistant germs in fish cultivation, is beneficial to cultivation environment protection and sustainable utilization, and lays a foundation for the future development of thebacteriophage serving as a novel bacterial infection controlling biological preparation.

Description

technical field [0001] The invention relates to a bacteriophage and its application in preparing medicines for preventing and / or treating diseases caused by Aeromonas hydrophila. Background technique [0002] Aeromonas ( Aeromonas ) is a genus in the Aeromonas family. It is a Gram-negative facultative anaerobic bacterium widely distributed in nature. According to their growth temperature and motility, the genus of bacteria can be divided into two categories: one is the mesophilic motile Aeromonas, including Aeromonas hydrophila ( Aeromonas hydrophila ), Aeromonas verkisii ( Aeromonas veronii ); the other is psychrophilic amotile Aeromonas such as Aeromonas salmonicida ( Aeromonassalmonicida ). Aeromonas hydrophila is widely distributed in various water bodies in nature, even in tap water. The bacteria can cause human diarrhea, food poisoning, trauma infection, etc., and severe cases can cause sepsis and peritonitis. It has been listed as the detection range of diarr...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N7/00A61K35/76A61P31/04C12R1/92
CPCA61K35/76A61P31/04C12N7/00C12N2795/00021
Inventor 肖炜张仕颖成亚辉钟莉瑶王永霞崔晓龙
Owner YUNNAN UNIV
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