A kind of bacteriophage and microecological agent for preventing and treating Aeromonas disease of aquatic animals
A probiotic, Aeromonas technology, applied in virus/phage, phage, medical raw materials derived from virus/phage, etc.
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Embodiment 1
[0017] Example 1 Screening and purification of phage 05A034
[0018] 1. Isolation of the pathogenic bacteria Aeromonas vermicularis in crucian carp large red gills
[0019] Cut the gill filaments of crucian carp suffering from big red gill disease in Jiangsu area in a sterile petri dish, rinse the surface of the gill filaments with 50 mL of 75% alcohol, place them in a sterile petri dish and chop them up and then dip the blood with an inoculation loop. Bacteria identification medium RYAN plate was streaked, cultured at 30°C for 24 hours, colonies were picked, streaked and purified on the same selection medium, and purified three times repeatedly, and the strains with the same morphology were picked with an inoculation loop to pick a single colony for inoculation. Put it into LB liquid medium, cultivate at 30°C for 12h, take 500 μL of bacterial liquid and 500 μL of 40% glycerol, mix and store in -80°C refrigerator.
[0020] 2. Expanded culture of Aeromonas wiertenii
[0021] ...
Embodiment 2
[0037] 1. Determination of optimal multiplicity of infection (MOI) of phage 05A034 against Aeromonas viridis
[0038]Take 8 100mL fresh LB liquid medium and process them separately. In treatment 1, add 1 mL at the same time to the content of 10 6 cfu / mL Aeromonas viridis liquid and 1mL content of 10 9 pfu / mL bacteriophage solution, add 1 mL at the same time for treatment 2, the content is 10 7 cfu / mL Aeromonas viridis liquid and 1mL content of 10 9 pfu / mL bacteriophage solution, add 1 mL of 10 8 cfu / mL Aeromonas viridis liquid and 1mL content of 10 9 pfu / mL bacteriophage solution, add 1 mL of 10 8 cfu / mL Aeromonas viridis liquid and 1mL content of 10 8 pfu / mL bacteriophage solution, add 1 mL of 10 8 cfu / mL Aeromonas viridis liquid and 1mL content of 10 7 pfu / mL bacteriophage liquid, add 1 mL of 10 8 cfu / mL Aeromonas viridis liquid and 1mL content of 10 6 pfu / mL bacteriophage liquid, add 1 mL of 10 8 cfu / mL Aeromonas viridis liquid and 1mL content of 10 5 pfu / mL bac...
Embodiment 3
[0050] The lysis ability and control effect of embodiment 3 phage 05A034
[0051] 1. Determination of the lysis ability of bacteriophage 05A034 on Aeromonas viridis
[0052] Phage 05A034 was counted and diluted to 1×10 8 , 1×10 7 , 1×10 6 , 1×10 5 , 1×10 4 5 concentrations of pfu / mL, with sterile water as the blank control, three parallels for each concentration. Take 6 tubes of cultured Aeromonas viridis bacterium solution, calculate the content of Aeromonas viridis by the dilution plate coating method before treatment, add 1 mL of 5 concentrations of phage solution and sterile water respectively, and then at 30 ℃ , After culturing for 12h under the condition of 150r / min, the lysis ability of phage 05A034 to Aeromonas viridis was determined by the dilution plate coating method to calculate the content of A. The results showed that bacteriophage 05A034 had better bactericidal effect on Aeromonas viridis, and the higher the concentration, the better the effect.
[0053] ...
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