Bacteriophage for preventing and treating aeromonas diseases of aquatic animals and application of bacteriophage
A technology of Aeromonas and aquatic animals, applied in the direction of viruses/bacteriophages, phages, medical raw materials derived from viruses/phages, etc.
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Embodiment 1
[0017] Example 1 Screening and purification of phage 05A034
[0018] 1. Isolation of Aeromonas victorii, the pathogenic bacterium of crucian carp red gill
[0019] Cut the gill filaments of crucian carp suffering from large red gill disease in Jiangsu area into a sterile petri dish, take 50mL of 75% alcohol to wash the surface of the gill filaments, put them in a sterile petri dish, chop them up, and then use an inoculation loop to dip in the blood. Streak on the RYAN plate of the bacterium identification medium, culture at 30°C for 24 hours, pick colonies, and purify by streaking on the same selection medium, repeat the purification 3 times, and use the inoculation loop to pick a single colony and inoculate the strains with the same shape. Put into LB liquid medium, culture 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. Expansion culture of Aeromonas victorii
[0021] For the expansion culture of...
Embodiment 2
[0037] 1. Determination of optimal multiplicity of infection (MOI) of phage 05A034 against Aeromonas verkistan
[0038]Take eight 100mL fresh LB liquid culture medium and process them separately. In treatment 1, add 1 mL at the same time with a content of 10 6 cfu / mL Aeromonas verdeii bacteria liquid and 1mL content is 10 9 pfu / mL phage liquid, add 1mL content to 10 at the same time for treatment 2 7 cfu / mL Aeromonas verdeii bacteria liquid and 1mL content is 10 9 pfu / mL phage liquid, add 1mL content to 10 at the same time for treatment 3 8 cfu / mL Aeromonas verdeii bacteria liquid and 1mL content is 10 9 pfu / mL phage liquid, add 1mL content to 10 at the same time for treatment 4 8 cfu / mL Aeromonas verdeii bacteria liquid and 1mL content is 10 8 pfu / mL phage liquid, add 1mL content to 10 at the same time for treatment 5 8 cfu / mL Aeromonas verdeii bacteria liquid and 1mL content is 10 7 pfu / mL phage liquid, add 1mL content to 10 at the same time for treatment 6 8 cfu / mL...
Embodiment 3
[0050] Example 3 Lysis ability and control effect of bacteriophage 05A034
[0051] 1. Determination of lysing ability of bacteriophage 05A034 on Aeromonas verkirea
[0052] The phage 05A034 was counted and diluted to 1×10 8 , 1×10 7 , 1×10 6 , 1×10 5 , 1×10 4 There are 5 concentrations of pfu / mL, and sterile water is used as the blank control, and three parallels are made for each concentration. Take 6 tubes of cultured Aeromonas verkirea bacteria liquid, calculate the content of Aeromonas verkirea by the dilution plate coating method before treatment, add 1mL phage liquid of 5 concentrations and sterile water respectively, and then incubate at 30°C After culturing for 12 hours under the condition of 150r / min, the lysing ability of phage 05A034 to Aeromonas verdeii was determined by calculating the content of Aeromonas verderichi after treatment by the dilution plate coating method. The results showed that bacteriophage 05A034 had a better bactericidal effect on Aeromona...
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