Pasteurella bacteriophage, bacteriophage composition and application of pasteurella bacteriophage
A Pasteurella and phage technology, applied in the field of microbiology, can solve the problem of no Pasteurella phage, etc., and achieve the effects of wide application, lower incidence probability, and lower cost
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[0033] The isolation culture and biological characteristic of embodiment 1 bacteriophage
[0034] (1) Recovery culture of strains and preparation of bacterial suspension
[0035] Pick the frozen bacteria solution of Pasteurella and draw a three-section line on the TSA plate (plus 5% newborn bovine serum), separate single colonies, and culture them in a 37°C incubator for 16-24 hours. Pick a single colony, inoculate in 5ml TSB broth (plus 5% newborn bovine serum), and incubate in an air shaker at 37°C at 180rpm for 18h to obtain a single bacterial suspension.
[0036] (2) Isolation and purification of phage
[0037] Take rabbit feces, litter, sewage and fur samples, add appropriate amount of TSB broth, shake at 37°C, 170rpm for 30min, centrifuge at 10000rpm for 5min, take the supernatant and filter it with a 0.22μm bacterial filter for later use.
[0038] Add each strain of Pasteurella in an amount of 80% in each Erlenmeyer flask, add the centrifuged supernatant (adding 5% ne...
Example Embodiment
[0055] The mensuration of embodiment 2 phage lysis spectrum and in vitro lysis test
[0056] (1) Determination of phage lysis profile
[0057] Adopt double-layer plate method to measure the cracking spectrum of phage, the steps are as follows: Obtain the bacterium suspension of host bacterium according to the method in embodiment 1 (47 strains of Pasteurella are respectively derived from dead rabbits in various regions of Shandong, Qinghai, Sichuan and other provinces. ). After incubating 300 μl of bacterial suspension and 300 μl of Pasteurella phage at 37°C for 5 minutes, add it to the upper layer of agar (with 5% newborn bovine serum) to prepare a double-layer plate, and place it in an incubator at 37°C for upside-down culture after the agar has solidified Observe the results overnight.
[0058] From the experimental results in Table 2, it is obtained that among 47 Pasteurella host bacteria from different sources, phage vB_PmuP_PS01 can lyse 40 of them, and the lysis rate ...
Example Embodiment
[0070] The safety test of embodiment 3 bacteriophage
[0071] Experimental method: Select 20 healthy BALB / C mice with a body weight of 18-20 g, half male and half male, and divide them into experimental group and control group, with half male and half female mice in each group, and the experimental group is given 200 μl of purified phage proliferation liquid ( Valence is 10 9PFU / ml), the control group was fed with normal saline (200 μl) for 7 days, the behavior of the mice was observed, and the changes of the organs of the mice were observed by autopsy.
[0072] Experimental results: The behavior of the mice in the experimental group and the control group showed no abnormalities. After the autopsy of the liver, lungs, heart, spleen, and kidneys, the organs were normal, and there was no significant difference in the test results between the experimental group and the control group.
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