大肠杆菌噬菌体的分离、生物学特性及初步应用  

Isolation,biological characteristics and application of Escherichia coli phage

作　　者：张明 寇文利 张国清 董浩 张永勤 宋钢 张德福 励建荣 ZHANG Ming;KOU Wenli;ZHANG Guoqing;DONG Hao;ZHANG Yongqin;SONG Gang;ZHANG Defu;LI Jianrong(College of Food Science and Technology,Institute of Ocean,Bohai University/Food Safety Key Lab of Liaoning Province/National and Local Joint Engineering Research Center of Storage,Processing and Safety Control Technology for Fresh Agricultural and Aquatic Products,Jinzhou 121013,China;Jinzhou Agricultural and Rural Comprehensive Service Center aquatic technology extension station,Jinzhou 121013,China;Shandong Meijia Group Co.,Ltd.,Rizhao 276806,China;Jinzhou Longhaixingang Science and Technology Co.,Ltd.,Jinzhou 121211,China;Linghai Dalian Seafood Process Co.,Ltd.,Jinzhou 121211,China)

机构地区：[1]渤海大学食品科学与工程学院,渤海大学海洋研究院,辽宁省食品安全重点实验室,生鲜农产品贮藏加工及安全控制技术国家地方联合工程研究中心,辽宁锦州121013 [2]锦州市农业农村综合服务中心水产技术推广站,辽宁锦州121013 [3]山东美佳集团有限公司,山东日照276800 [4]锦州市龙海馨港科技有限公司,辽宁锦州121211 [5]凌海市达莲海珍品养殖有限责任公司,辽宁锦州121211

出　　处：《中国渔业质量与标准》2023年第5期31-38,共8页Chinese Fishery Quality and Standards

基　　金：国家自然科学基金(31801662);国家重点研发计划“蓝色粮仓科技创新”重点专项(2019YFD0901702)。

摘　　要：本研究以大肠杆菌SPHJ965为宿主菌,从生活污水中分离到一株大肠杆菌噬菌体,并命名为ΦSPHJ965,进一步分析了该噬菌体对温度和pH的稳定性、对紫外线和氯仿的敏感性,测定了最佳感染复数及一步生长曲线,最后进行了清除污水中大肠杆菌的初步应用。结果表明,ΦSPHJ965的效价为3.4×10^(9)PFU/mL,对pH以及氯仿的耐受性较强,但对温度和紫外线较敏感,60℃以上时滴度有明显的下降,经紫外线照射30 min后便丧失对宿主菌的裂解能力。该噬菌体最佳感染复数是0.00001,潜伏期为10 min,暴发期为90 min,暴发量为40 PFU/cell。对宿主菌的清除实验表明,ΦSPHJ965在6 h内对宿主菌有较好的抑制作用,6 h后实验组的OD_(595)值逐渐升高,但始终远低于对照组值,说明该噬菌体对宿主菌具有一定的裂解效果,但不能将宿主菌完全裂解。污水经ΦSPHJ965处理1 h后菌落总数和大肠杆菌数均有明显的下降,说明该噬菌体有较好的抑菌效果。综上可知,ΦSPHJ965的效价较高,对大肠杆菌的去除效果较好,具有作为大肠杆菌生物清除剂的应用潜能。A bacteriophage of E.coli SPHJ965 was isolated from domestic sewage and named as ΦSPHJ965.The stability of the bacteriophage to temperature and pH,the sensitivity to UV and chloroform were further analyzed.The optimal multiplicity of infection and one-step growth curve were determined.Finally,the preliminary application of E.coli removal was carried out.The results showed that the titer of ΦSPHJ965 was 3.4×10^(9) PFU/mL,and it had strong tolerance to pH and chloroform,but it was sensitive to temperature and UV.The titer decreased significantly when incubated above 60℃.After UV irradiation for 30 min,it lost its lytic ability to host bacteria.The optimal multiplicity of infection was 0.00001,The phage demonstrated the incubation period was 10 min,burst period was 90 min and burst amount was 40 PFU/cell.The antibacterial experiment showed that ΦSPHJ965 had a good inhibitory effect on the host bacteria within 6 h.After 6 h,the OD_(595) value of the experimental group increased gradually,but remained far lower than that of the control group,indicating that the phage had a good lytic effect on the host bacteria,but it could not completely lyse.After the wastewater was treated with ΦSPHJ965 for 1 h,the total number of colonies and Escherichia coli decreased significantly,indicating that the phage had good antibacterial effect.In conclusion,ΦSPHJ965 has high titer,strong tolerance to temperature and pH,and good removal effect on Escherichia coli,which has potential application as a biological scavenger of Escherichia coli.

关 键 词：大肠杆菌 噬菌体 分离 生物学特性 

分 类 号：Q93-3[生物学—微生物学]