鸡源大肠杆菌噬菌体的分离鉴定及组合应用  

Isolation,identification and combined application of phages of Escherichia coli from chicken

作　　者：路建彪[1] 王顺山 王俊丽 李玉保[1] 司振书[1] 杨文文 庞喆羽 徐欣璐 管玉堂 孟凡达 丁宁宁 LU Jianbiao;WANG Shunshan;WANG Junli;LI Yubao;SI Zhenshu;YANG Wenwen;PANG Zheyu;XU Xinlu;GUAN Yutang;MENG Fanda;DING Ningning(College of Agronomy,Liaocheng University,Liaocheng 252059,China;Beijing Huaxia Xingyang Biological Technology Co.,Ltd.,Beijing 102600,China;Yanggu County Agricultural Integrated Service Center,Yanggu 252025,China)

机构地区：[1]聊城大学农学院,山东聊城252059 [2]北京华夏兴洋生物科技有限公司,北京102600 [3]阳谷县农业综合服务中心,山东阳谷252025

出　　处：《黑龙江畜牧兽医》2021年第20期61-65,69,154,共7页Heilongjiang Animal Science And veterinary Medicine

基　　金：山东省重点研发计划项目(2019QYTPY011,2019GNC106082);山东省自然科学基金项目(ZR2020MC175);聊城大学实验技术项目(LDSY2014054)。

摘　　要：为探究大肠杆菌噬菌体的生物学特性及其混合制剂的应用效果,试验从山东地区分离纯化鸡源大肠杆菌及大肠杆菌噬菌体,对噬菌斑形态和大小差异较大的噬菌体进行了生物学特性(噬菌体效价、最适温度、最适pH值、最佳感染复数、一步生长曲线、最佳保存温度)测定,并测定了部分噬菌体的宿主谱,选取裂解率较高的10株噬菌体组合成噬菌体混合制剂,测定其对大肠杆菌(实验室保存的100株大肠杆菌、全国不同地区养鸡场分离的100株大肠杆菌)的裂解率。结果表明:从山东地区分离并纯化鸡源大肠杆菌195株,大肠杆菌噬菌体155株。挑取两株噬菌斑形态和大小差异较大的噬菌体(分别命名为PLCF1和PLCF72)进行了生物学特性测定,噬菌体PLCF1和PLCF72效价分别为4.5×10^(10) pfu/mL、1.5×10^(9) pfu/mL,在温度40℃以内可以保持较高的活性,最佳pH值为6~8,噬菌体PLCF1和PLCF72最佳感染复数分别为0.001,0.01;一步生长曲线显示噬菌体PLCF1的裂解性能强于噬菌体PLCF72;噬菌体适宜保存温度为-20℃或-80℃。选取83株噬菌体对282株大肠杆菌菌株进行裂解,裂解率可达78.01%,混合制剂对实验室保存的100株大肠杆菌的裂解率为62.00%,对全国不同地区采集的100株大肠杆菌的裂解率为41.00%。说明两株噬菌体生物学特性存在一定的差异,噬菌体混合制剂对不同来源的大肠杆菌裂解率不同。In order to explore the biological characteristics of Escherichia coli(E.coli) phages and the application effect of their mixed preparation, this experiment isolated and purified chicken-derived E.coli and E.coli phage from Shandong Province;the biological characteristics of phages with large differences in plaque morphology and size were determined(phage titer, optimal temperature, optimal pH value, optimal multiplicity of infection, one-step growth curve, optimal storage temperature),and determined the host spectrum of some phage;the 10 strains phage with higher lysis rate was selected to combine into a phage mixed preparation which was determined for its lysis rate to E.coli(100 strains of E.coli stored in the laboratory, and 100 strains of E.coli isolated from chicken farms in different regions of China). The results showed that 195 strains of E.coli from chicken and 155 strains of E.coli phages from Shandong Province were isolated and purified. Two phages(named PLCF1 and PLCF72) with large differences in plaque morphology and size were selected for biological characterization. The phage titers of PLCF1 and PLCF72 was 4.5×10^(10) pfu/mL,1.5×10^(9) pfu/mL,respectively, and could maintain high activity within 40 ℃,and the optimal pH of 6-8;the optimal multiplicity of infection for PLCF1 and PLCF72 were 0.001 and 0.01 respectively;the one-step growth curve showed that the lysis performance of PLCF1 was stronger than that of PLCF72;the optimal storage temperature for phage was-20 ℃ or-80 ℃. Eigty-three bacteriophages were selected to lyse 282 E.coli strains, and the lysis rate could reach 78.01%. The lysis rate of the mixed preparation on 100 strains of E.coli stored in the laboratory was 62.00%,and the lysis rate on 100 strains of E.coli collected from different regions of China was 41.00%. The results suggested that there were certain differences in the biological characteristics of the two phage strains, and the bacteriophage mixed preparation had different lysis rates for E.coli from different sources

关 键 词：大肠杆菌 噬菌体 生物学特性 噬菌体混合制剂 裂解率 

分 类 号：S859.799.9[农业科学—临床兽医学]