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Novel vibrio alginolyticus bacteriophage and composition and application thereof

A technology of Vibrio alginolyticus and phage, which is applied in the field of microorganisms, can solve the problem of no bacteriophage of Vibrio alginolyticus, and achieve the effects of no toxic side effects, high safety, and cost reduction

Active Publication Date: 2020-09-29
QINGDAO PHAGEPHARM BIO TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented technology describes an improved way for killing harmful microorganisms called VBS (Vibronchoa fragilis) without causing any negative reactions like other organism's. These new methods use specific types of virus known as VBLV(vibrate brittlenbergii), BLVBNZM01, PDB06039, LAVGINAVERE1, GALYX-3, YELF4-5, XEN-4115, TMB-26-1, HIV-1, EBOKO7-8, KSHV). By adding these viruses to fish feed containing them during their growth cycle, they may stop disease outbreaks while still maintaining effective control over the remaining population of infectious seafood species such as shrimp farming.

Problems solved by technology

This patents discusses different methods currently available but none effectively prevents or controls VLBI virus in cultured salmon farming systems due to its ability to resist chemical substances like chloroquinetisulfate sodium sulfuricate solution commonly known as vibratozones. Current solutions involve adding these compounds into feedstuffs along with certain types of seafood ingredients. These techniques result in increased risk of developing drug-related illnesses associated with VLBIOVlAIDOXIS type BVDHYE, making them challenged against international trade standards.

Method used

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  • Novel vibrio alginolyticus bacteriophage and composition and application thereof
  • Novel vibrio alginolyticus bacteriophage and composition and application thereof
  • Novel vibrio alginolyticus bacteriophage and composition and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Example 1: Isolation, purification and amplification of phage

[0039] Sewage samples from the Chengyang Seafood Wholesale Market in Qingdao City, Shandong Province were taken, added to Vibrio alginolyticus logarithmic phase bacterial liquid J20, and an appropriate amount of 2216E medium, and cultured on a shaker at 37°C and 170rpm for 16h. Take out 5ml of the liquid, centrifuge at 11000rpm / min for 10min, and filter the supernatant through a 0.22μm sterile microporous membrane to obtain the phage stock solution, which is stored at 4°C.

[0040] The phage was isolated by the double-layer plate method, and the phage stock solution was diluted 10 times, and 100 μl of the diluted solution was mixed with 200 μl of the corresponding Vibrio alginolyticus bacterial solution, incubated at 37° C. for 5 minutes, and mixed into the upper layer of agar (agar concentration: 0.8%). Spread evenly on the lower layer of 2216E agar (1.5% agar concentration) plates, cool and incubate overn...

Embodiment 2

[0043] Electron microscope observation of embodiment 2 phage

[0044] The phage suspension isolated above was dropped on a copper grid covered with polyvinyl formaldehyde film, stained with 2% phosphotungstic acid for 5-10 minutes, and observed with a Hitachi HT7700 transmission electron microscope after drying.

[0045] Observed under the transmission electron microscope, its shape is as figure 2 As shown, the phage has an icosahedral head with a short but not contractile tail, the head diameter is about 54nm, and the tail length is about 15nm; according to the classification method of the International Committee on Taxonomy of Viruses (ICTV), this application The phage morphology conforms to the characteristics of the Brachyphage family, belongs to the Brachyphage, and is named vB_ValP_PJ32.

Embodiment 3

[0046] Genome Analysis of Example 3 Phage

[0047] Extract the genome of Pseudomonas aeruginosa phage vB_ValP_PJ32, and carry out whole genome sequencing, obtain:

[0048] The full genome length of phage vB_ValP_PJ32 is: 80293bp; among them, the tail protein gene sequence related to phage host recognition is shown in sequence 1 in the sequence table, and the highly conserved terminal enzyme large subunit (terminase large subunit) gene sequence is shown in sequence For sequence 2 in the list, see sequence 3 in the sequence listing for the sequence of DNA polymerase (DNA polymerase) gene. The specific information of the above genes is shown in Table 1 below.

[0049] Using BLAST online tool (http: / / blast.ncbi.nlm.nih.gov / ) to perform sequence similarity comparison analysis on phage genomes. The phage with the highest homology was Vibrio phage Ares1, and the homology between phage vB_ValP_PJ32 and this phage only reached 76.22%. The above results indicated that phage vB_ValP_P...

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Abstract

The invention discloses a novel vibrio alginolyticus bacteriophage and a composition and an application thereof. The vibrio alginolyticus bacteriophage vB _ ValP _ PJ32 has been preserved in China General Microbiological Culture Collection Center on November 4, 2019, and the preservation number is CGMCC No. 18860. The novel vibrio alginolyticus bacteriophage not only has a wider lysis spectrum, but also has excellent acid-base tolerance, can adapt to a harsher application environment, and is used for preventing and treating vibrio alginolyticus infection. The bacteriophage and the compositioncompounded by using the bacteriophage can be used for preparing a medicine for preventing and treating diseases caused by vibrio alginolyticus infection in aquaculture, a prawn feed additive, an aquaculture farm environment disinfectant, a detection test kit and a biological bacteriostatic agent for disinfection of fresh food; the composition can be used for various links which are easily lost dueto vibrio alginolyticus infection in the aquaculture process, daily disinfection of the aquaculture market and the aquaculture environment and the like, and is beneficial to healthy development of the aquaculture industry.

Description

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Claims

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Application Information

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Owner QINGDAO PHAGEPHARM BIO TECH CO LTD
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