A strain of Vibrio harveyi phage, its phage composition and its application
A technology of Vibrio harveyi and bacteriophage, applied in the field of microorganisms, can solve problems such as antibiotic residues, achieve the effects of easy industrial production, reduce costs, and reduce the probability of disease occurrence
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Problems solved by technology
Method used
Image
Examples
Embodiment 1
[0032] The isolation and cultivation of embodiment 1 phage
[0033] (1) Preparation of host bacterial suspension:
[0034] The host bacterium K005 was divided and marked on the TCBS plate, and a single colony was picked and inoculated into 5ml 2216E liquid medium, and multiplied to the logarithmic growth phase at 170rpm and 37°C to obtain a fresh bacterial solution.
[0035] (2) Isolation and purification of phage
[0036] (1) Isolation of phage
[0037] Sewage collected from a seafood market in Qingdao City, Shandong Province was centrifuged at 11,000 r / min for 5 minutes, and the supernatant obtained by centrifugation was filtered with a 0.22 μm filter membrane, then added with 2216E medium and 100 μl of host bacteria suspension, and the mixture was heated at 170 rpm 1. After culturing at 37°C for 16 hours, centrifuge at 11,000 r / min for 5 minutes, and finally filter with a 0.22 μm filter membrane to obtain the phage stock solution.
[0038] The phage stock solution was di...
Embodiment 2
[0046] The biological characteristic determination of embodiment 2 bacteriophage
[0047] (1) Electron microscope detection of phage
[0048] Take 20μl for a titer of 10 10 The phage of PFU / ml was dropped on the copper grid, precipitated for 15 minutes, absorbed the excess liquid with filter paper, stained with 2% phosphotungstic acid for 30 minutes, and observed with electron microscope after drying.
[0049] The result is as figure 1 As shown, the length of the phage head is 80nm, the head width is 75nm, and the retractable tail length is 125nm. According to the classification standard identification of the ninth report of the International Committee on Taxonomy of Viruses (ICTV), it can be It was determined that the phage belonged to the family Myocaudidae, and it was named vB_KaS_PK08.
[0050] (2) Determination of lysis profile of phage
[0051] Experimental method: In this example, the double-layer plate method was used to measure the bacteriophage lysis spectrum. Ta...
Embodiment 3
[0073] Embodiment 3 phage lysis experiment in vitro
[0074] The phage suspension and the host bacteria were added to 100ml 2216E liquid medium according to the ratio of MOI0.01, and the final concentration of clinically isolated pathogenic Vibrio harveyi strain KO11 was 5.00×10 6 CFU / ml, the final concentration of phage was 5.00×10 4 PFU / ml, add the same amount of sterile 2216E medium as the phage solution to the control group, mix the bacteriophage and phage, shake and culture in a shaker at 37°C and 170rpm, measure the OD value at regular intervals until the mixture becomes clear .
[0075] The result is as Figure 5 Shown: the phage can effectively inhibit the host bacteria, OD after 2h 600 Significantly decreased, indicating that the bacterial concentration decreased sharply, and the OD at 3h 600 has dropped to 0.3, and at this time the OD of the host bacteria in the control group 600 The value has risen to more than 1.0, and the mixed solution added to the phage sus...
PUM
Login to View More Abstract
Description
Claims
Application Information
Login to View More