1株貂源绿脓杆菌噬菌体的分离与鉴定

Isolation and Identification of Pseudomonas aeruginosa Phage from Mink Feces

从水貂粪便中分离并纯化1株绿脓杆菌噬菌体,为测定其各项生物学特性。从水貂场采集粪便,利用双层平板法分离噬菌体,对分离到的噬菌体进行电镜形态观察、裂解谱、效价、最佳感染复数、温度和pH值敏感性以及一步生长曲线的测定。结果显示,从水貂样品中分离纯化得到一株绿脓杆菌噬菌体,命名为phage30。电镜形态特征表明,该噬菌体头部为二十面体的立体对称结构,直径约为60nm,尾部长180nm,属于有尾噬菌体目,长尾噬菌体科;噬菌体裂解率为29.63%(8/27);在宿主菌中增殖的效价为4.3×10^10PFU/mL。生物学特性测定结果表明,噬菌体在pH值5~11、温度40℃以下裂解活性基本保持不变,最佳感染复数为0.01;一步生长曲线测定结果显示,噬菌体潜伏期约为60min,暴发期约120min,裂解量约为156。表明从水貂粪便分离到这1株绿脓杆菌噬菌体,属长尾噬菌体科,具有宿主谱宽、裂解性能高、pH值和热稳定性好的特征,具有潜在的开发价值。

In order to isolated and purified a phage of P. aeruginosa from mink feces and determined its biological properties. P. aeruginosa were used as host cell to isolate phages from mink feces by double layer plate method. The morphology, host range, potency, the optimal multiplicity of infection (MOI), one-step growth curve and the stability to temperature and pH character were lanalyzd.(Result) A phage strain of P. aeruginosa named as phage30 was isolated and purified. Morphological features observed using Electron microscopy indicated that phage30 belonged to the Siphoviridae in Caudovirales, which consists of an icosahedral head with a diameter of about 60 nm and a long tail with an approximate length of 180 nm. Phage lysis rate was 29. 63%(8/27). The phage titer was 4. 3 ×10^10 pfu·mL ^-1 . The biological characteristic results showed that the phage titer kept in the same level in the range of pH 5 ~11 or at 40 ℃, and the MOI was 0. 01. One-step growth curve results indicated that the latent period was about 60 min, the burst period was about 120 min with a burst size of 156. A P. aeruginosa phage30 was isolate from mink feces, which belongs to Siphoviridae in Caudovirales. Phage30 shows a wide host range, a strong lysis and a stable ability in wide temperatures and pH range showing a potential development value.