FSN17-1 Photobacterium damselae phage FSN17-1 and application

A technology of photobacteria and mermaids, applied in the field of bioengineering, can solve the problems of narrow cleavage spectrum, etc., and achieve the effects of fast proliferation, high application value and commercial value, and convenient fermentation and large-scale production

Active Publication Date: 2019-04-23
JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The research on the phage of P. mermaidus is limited. At present, only Yamaki, S, etc. in Japan have isolated a phage of P. One of them had a cleavage rate of only 10% (Yamaki, S, Kawai, Y., & Yamazaki, K. Characterization of a novel bact

Method used

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  • FSN17-1 Photobacterium damselae phage FSN17-1 and application
  • FSN17-1 Photobacterium damselae phage FSN17-1 and application
  • FSN17-1 Photobacterium damselae phage FSN17-1 and application

Examples

Experimental program
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Example Embodiment

[0035] Example 1 Isolation and preparation of phage

[0036] The host bacteria of the experimental phage of the present invention is Photobacterium mermaid clinical isolate NDA12 (GenBank accession number is MK285669).

[0037] The sample of the present invention is the abdominal cavity content of diseased large yellow croaker collected from Sanduao, Ningde City, Fujian Province. The sample is cut into pieces and dissolved in 15 mL SM liquid, centrifuged at 12000 rpm for 20 minutes, and then filtered with a 0.22 μm filter membrane. Take 10mL filtered supernatant, add 0.5mL host bacteria overnight culture, then add sterile CaCl 2 Mix the mother liquor to a final concentration of 1.25mM, add 20mL of 2216E medium, place it in a 30℃ incubator for 6-8h, take the above culture at 12000rpm, centrifuge for 30min, take the supernatant; take this supernatant 10mL, Add 0.5mL host bacteria overnight culture, add sterile CaCl 2 Mix the mother liquor to a final concentration of 1.25mM, add 20mL ...

Example Embodiment

[0040] Example 2 Amplification and purification of phage

[0041] On the double-layer plate forming the plaque in Example 1, use the pipette tip to pick a single plaque with a larger diameter, inoculate it in 5mL 2216E medium, and add the host bacterium solution in the logarithmic growth phase 0.1mL, mix well, incubate at room temperature for 15 minutes, incubate at 30°C for 12 hours, centrifuge at 12000 rpm, 4°C for 10 minutes, take the supernatant, and add 0.3% chloroform. Pick a single plaque 4-5 times repeatedly until the phage is purified into plaques of the same size.

[0042] Take 1 mL of freshly cultured host bacteria and add 0.5 mL of phage lysate (a single phage culture and host bacteria are in a ratio of 1:1, 1:10, and 1:100 respectively). Incubate at 30°C for 20 minutes to make the phage particles adsorb to the host bacteria; add 100mL 2216E liquid medium, and then add CaCl 2 The mother liquor was cultured to a final concentration of 1.25 mM, shake culture at 30°C for ...

Example Embodiment

[0047] Example 3 The influence of temperature and pH on phage

[0048] Take 0.1mL 1×10 9 Pfu / mL purified phage FSN17-1 (provided in Example 2) was placed in a water bath at 30-90°C for 30 min or 60 min, and the sample was cooled and its titer was measured; respectively, peptone water with pH 3.0-12.0 and 1×10 8 The pfu / mL purified phage was mixed in equal amounts, and the titer was measured after 2 hours at 30°C in a water bath.

[0049] The effect of temperature on the survival of bacteriophages is as image 3 Shown by image 3 It can be seen that there is no significant change in the activity of the phage after 30min or 60min at 30-60℃; after the action at 70℃, the titer of the phage drops significantly; after the action at 80℃ for 1h, no phage survives.

[0050] The effect of pH on phage survival is as Figure 4 Shown by Figure 4 It can be seen that at pH 3.0, no viable phages can be detected; at pH 4.0 and 11.0, the phages are still alive, but their titer is relatively low; at pH...

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Abstract

The invention discloses an FSN17-1 Photobacterium damselae phage FSN17-1 and application reserved in China Center for Type Culture collection, and the preservation number is CCTCC NO: M2018393; the FSN17-1 Photobacterium damselae phage can stably survive under the condition that the temperature is 30-60 DEG C and the pH is 5.0-9.0, the hydrophily is good, the splitting spectrum is wide, and the FSN17-1 Photobacterium damselae phage has the good splitting capability for Photobacterium damselae, has the good treatment effect on infection of Photobacterium damselae, and a novel way is provided for pathogenic bacterium prevention and control.

Description

technical field [0001] The invention belongs to the technical field of bioengineering, and in particular relates to a phage strain of Photobacterium mermaidus and its application in preventing and treating the infection of Photobacteria mermaidus. Background technique [0002] Photobacterium damselae, a Gram-negative bacterium, is the causative bacterium of Photobacterium damselae. Photobacterium mermaidi widely exists in the marine environment, can infect a variety of marine economic fish, and has no obvious host specificity. The typical characteristic of sick animals is hemorrhagic sepsis, with rapid onset and high mortality, often causing major economic losses. loss. In addition, the pathogen can also infect humans and mammals, posing a threat to human life and health. [0003] As early as 1963, Photobacterium mermaid was found in natural populations of white bass and striped bass in the Chesapeake coast of the United States. Since 1990, outbreaks of this pathogen have ...

Claims

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Application Information

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IPC IPC(8): C12N7/00A61K35/76A61P31/04A23K10/18A23K50/80A01N63/00A01P1/00C12R1/92
CPCA61P31/04A01N63/00A23K10/18A23K50/80A61K35/76C12N7/00
Inventor 庞茂达王冉张辉孙利厂包红朵吴立婷
Owner JIANGSU ACADEMY OF AGRICULTURAL SCIENCES
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