A kind of broad-spectrum Salmonella phage biological bactericide and its application
A technology of phage organisms and Salmonella, applied in the field of bioengineering, can solve problems such as multi-drug resistance, hinder the development of poultry industry, threaten the health of humans and animals, and achieve the effect of low cost and wide source
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Embodiment 1
[0023] Example 1 Isolation, amplification and purification of Salmonella phage
[0024] The fecal and urine sewage samples from the chicken farm in this example came from a chicken farm in Nantong City, Jiangsu Province; the host bacterium was Salmonella Enteritidis SM-54-KDE preserved in this laboratory, which was isolated from sick chickens in a chicken farm in Jinan City.
[0025] Phage isolation: Collect fecal and urine sewage samples from chicken farms, filter them through double-layer filter paper, centrifuge at 12000×g for 20 minutes, and then filter the supernatant with 0.45 μm and 0.22 μm filter membranes respectively; take 10 mL of the filtrate, add 0.5 mL Overnight cultured host bacteria (with a bacterial content of 10 9 cfu / mL), then add 20mL 4×LB medium, place it at 37°C and incubate for 12-16h, the next day, take the above culture and centrifuge at 12000×g for 10min, take the supernatant and add 0.3% of the volume of the supernatant in chloroform Standby; use sp...
Embodiment 2
[0030] Example 2 Isolation, amplification and purification of Salmonella typhimurium phage
[0031] The feces and urine sewage samples from chicken farms in this example came from a chicken farm in Nantong City, Jiangsu Province; the host bacteria was Salmonella typhimurium HNER178, which was isolated from a sick and dead chicken in a farm in Jiangxi Province, and was preserved in our laboratory.
[0032] The isolation, amplification and purification of Salmonella typhimurium phage, except that the host bacteria is Salmonella typhimurium HNER178, the rest of the steps are the same as in Example 1, and the titer of the purified Salmonella typhimurium phage is ≥10 8 pfu / mL.
[0033] The applicant self-named the Salmonella typhimurium phage as JX178, and its microbiological characteristics are as follows: figure 2As shown, the phage has a polyhedral head structure and a long tail, and should belong to the longtail virus family, and the phage belongs to the lytic phage.
[0034...
Embodiment 3
[0035] Example 3 Isolation, Amplification and Purification of Salmonella Delphi Phage
[0036] In this example, the sewage sample came from a farmer's market in Jiangsu Province; the host bacteria was Salmonella delbei HNER067-2, which was isolated from a farmer's market in Jiangxi Province and preserved in our laboratory.
[0037] For the isolation, amplification and purification of Salmonella Delphi phage, except that the host bacterium is Salmonella Delphi, the other operating steps are the same as those in Example 1, and the titer of the purified Salmonella Delphi phage is ≥10 8 pfu / mL. The applicant self-named the Salmonella phage JX067-2, and its microbiological characteristics are as follows: image 3 As shown, the phage has a polyhedral head structure and a long tail, and should belong to the longtail virus family, and the phage belongs to the lytic phage.
[0038] The applicant deposited the purified Salmonella delphi phage JX067-2 in the China Center for Type Cultu...
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