Salmonella bacteriophage resistant to antiviral drugs and application of salmonella bacteriophage
An antiviral drug and Salmonella technology, applied to viruses/phages, medical raw materials derived from viruses/phages, phages, etc., can solve the problems of bacterial resistance, untimely treatment, and ineffective medication, etc. Good tolerance and titer stability
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Embodiment 1
[0033] Embodiment 1 Isolation and identification of pathogenic bacteria and bacteriophage
[0034] 1. Isolation and identification of pathogenic Salmonella BS20133
[0035] Sampling from the diseased farm, aseptically take the liver of diseased poultry, streak it on the selective medium (SS agar), culture it at 37°C for 18-24 hours, and form a round, flat, neat edge and smooth surface on the medium Wet black colonies, pick typical colonies and continue to streak and purify 3 times, then pick a single colony and inoculate in 5mL LB broth, shake and culture at 200rpm at 37°C for 8h to obtain a uniform turbid bacterial suspension. Then through 16sRNA molecular identification and serotype identification, it was determined to be pathogenic Salmonella, and one of the strains was named BS20133 and stored in a -80°C refrigerator.
[0036] 2. Isolation and identification of Salmonella phage RDP-SA-20032
[0037] (1) Manure treatment: Take manure from the farm, weigh 5g of chicken man...
Embodiment 2
[0043] Morphological observation of embodiment 2 bacteriophage
[0044] Using phosphotungstic acid negative staining method: drop phage liquid (titer 10 10 pfu / m L) 100μL, put the film side of the copper mesh on the phage droplet, take it off after 10min, let it dry naturally in the air for 2-3min, then drop a drop of 2% phosphotungstic acid (PTA) on the copper mesh The aqueous solution was stained, removed after 10 minutes, dried in the air for 10-15 minutes, observed with an electron microscope, and a clear phage image was selected for photographing. photo by electron microscope figure 1 It can be seen that the bacteriophage has a polyhedral stereosymmetric head, wrapped in nucleic acid, about 65-40nm in diameter, has a tail about 100nm long, has a tail sheath, and the neck is connected to the head and tail. Classified as Tauroviridae.
Embodiment 3
[0045] The mensuration of embodiment 3 phage thermostability
[0046] Dispense the phage stock solution into 50 ml sterilized Erlenmeyer flasks, incubate at 0°C, 30°C, 40°C, 50°C, 60°C, 70°C, 80°C and 90°C for 30min and 60min respectively, and then use After 10-fold serial dilution of normal saline, spread double plates to determine its titer; the results are as follows: figure 2 shown.
[0047] Depend on figure 2 It can be seen that the titer of bacteriophage RDP-SA-20032 is stable within 60°C and remains at 10 11 About pfu / mL; when treated at 60°C for 30min and 60min, the phage titer only decreased by 1 order of magnitude; when incubated at 80°C for 30min and 60min, it was still active, indicating that the phage has good high temperature resistance.
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