[1]李陇平,杨吉,白蹉蹉,等.一株绒山羊源大肠杆菌噬菌体φPTK的分离鉴定及生物学特性分析[J].江苏农业学报,2018,(04):847-853.[doi:doi:10.3969/j.issn.1000-4440.2018.04.019]
　LI Long-ping,YANG Ji,BAI Cuo-cuo,et al.Isolation, identification and biological properties of a lytic phage against goat (Capra hircus)-associated Escherichia coli[J].,2018,(04):847-853.[doi:doi:10.3969/j.issn.1000-4440.2018.04.019]
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一株绒山羊源大肠杆菌噬菌体φPTK的分离鉴定及生物学特性分析()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2018年04期
页码:
847-853
栏目:
畜牧兽医·水产养殖
出版日期:
2018-08-25

文章信息/Info

Title:
Isolation, identification and biological properties of a lytic phage against goat (Capra hircus)-associated Escherichia coli
作者:
李陇平; 杨吉; 白蹉蹉; 杨悦; 屈雷
（榆林学院陕西省陕北绒山羊工程技术研究中心，陕西榆林719000）
Author(s):
LI Long-ping; YANG Ji; BAI Cuo-cuo; YANG Yue; QU Lei
(Shaanxi Provincial Engineering and Technology Research Center of Cashmere Goats, Yulin University, Yulin 719000, China)
关键词:
绒山羊; 羔羊痢疾; 大肠杆菌; 噬菌体; 生物学特性
Keywords:
cashmere goat; diarrhoea; Escherichia coli; bacteriophage; biological properties
分类号:
S858.27
DOI:
doi:10.3969/j.issn.1000-4440.2018.04.019
文献标志码:
A
摘要:
以从患羔羊痢疾的羔羊粪便中分离的1株大肠杆菌为宿主菌，从污水中分离纯化1株烈性大肠杆菌特异性噬菌体φPTK，并对其生物学特性进行研究，为研制一种替代抗生素的有效生物防治方法提供资源。双层琼脂平板培养后，噬菌斑呈现清晰透明、边缘光滑的圆形，直径 2～3 mm。噬菌体φPTK生物学特性试验结果表明：φPTK对大肠杆菌的最佳感染复数为0.000 1；φPTK吸附和感染宿主菌的潜伏期为15 min，裂解期约195 min，裂解量93；噬菌体浓缩液经SDS-PAGE分析可以观察到至少6条蛋白质条带，相对分子质量为 2.5×104～1.8×105，说明其蛋白质外壳至少含有6个结构蛋白；在 30～90 ℃范围内φPTK的裂解活性均较强，30～40 ℃时活性最高，90 ℃处理90 min后仍然具有较强的活性；对酸碱的耐受力较强，pH值在5至9范围内均有较强的裂解活性，当pH值超过9后活性较低；对紫外线的耐受能力较强，经紫外线照射8 h后仍然具有较强的裂解能力。这株烈性大肠杆菌特异性噬菌体φPTK最佳感染复数较小，吸附和感染宿主菌的潜伏期短，裂解能力强，杀菌效果好，有较强的环境适应能力，具有研发成为有效防治羔羊大肠杆菌病的生物杀菌制剂的潜力。
Abstract:
In order to provide an alternative antibiotics for the prevention of lamb dysentery, a lamb-associated Escherichia coli virulent specific bacteriophage, named φPTK, was isolated and purified from sewage, and its biological characteristics were determined. The plaque of the phage φPTK showed circular, clear and transparent with smooth edge, showing 2-3 mm of diameter, using the double-layer agar plate method. The analysis results of its biological properties showed that the optimal multiplicity of infection (MOI) was 0.000 1. The determination of one-step growth kinetics showed a short latency period of about 15 min and a burst phase time of 195 min and a relatively larger burst size was 93. The structural proteome of phage φPTK was analyzed by SDS-PAGE, at least six protein bands could be observed with a relative molecular mass of 2.5×104-1.8×105, indicating that the phage φPTK had at least six structural proteins. In the range of 30-90 ℃, φPTK had a strong lytic activity, and the activity was highest at 30-40 ℃. After treated 90 minutes at 90 ℃, it still had a strong activity. Phage φPTK was quite stable in a wide range of pH values (pH 5-9). Moreover, phage φPTK was resistant to ultraviolet ray treatment, and the titer was still remained a high level after ultraviolet irradiation for eight hours. In the present study, an E. coli-specific virulent phage was isolated, the phage φPTK had a short latency period, strong lytic activity and ability of sterilization. It has the potential to be developed as a new approach to control the lamb-associated E. coli disease.

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备注/Memo

备注/Memo:
收稿日期：2017-12-27 基金项目：陕西省高校科协青年人才托举计划项目（20160236）；榆林学院高层次人才科研启动基金项目（16GK21） 作者简介：李陇平（1985-），男，陕西陇县人，博士，讲师，主要从事畜牧微生物研究。(E-mail)llp_315@163.com
更新日期/Last Update: 2018-09-04