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Isolation and characterization of a novel Escherichia coli O157:H7-specific phage as a biocontrol agent.
Journal of Environmental Health Science and Engineering ( IF 3.4 ) Pub Date : 2020-02-06 , DOI: 10.1007/s40201-020-00452-5
Cheonghoon Lee 1, 2 , In Young Choi 3 , Do Hyeon Park 3 , Mi-Kyung Park 3, 4
Affiliation  

Purpose

Escherichia coli O157:H7 is one of the major foodborne pathogens of global public concern. Bacteriophages (phages) have emerged as a promising alternative to antibiotics for controlling pathogenic bacteria. Here, a lytic E. coli O157:H7-specific phage (KFS-EC) was isolated, identified, and characterized to evaluate its potential as a biocontrol agent for E. coli O157:H7.

Methods

KFS-EC was isolated from slaughterhouse in Korea. Morphological analysis, genomic analysis and several physiological tests were performed to identify and characterize the KFS-EC.

Results

A specificity test indicated KFS-EC was strictly specific to E. coli O157:H7 strains among 60 bacterial strains tested. Morphological and phylogenetic analyses confirmed that KFS-EC belongs to the Rb49virus genus, Tevenvirinae subfamily, and the Myoviridae family of phages. KFS-EC genome consists of 164,725 bp and a total of 270 coding sequence features, of which 114 open reading frames (ORFs) were identified as phage functional genes. KFS-EC does not contain genes encoding lysogenic property and pathogenicity, which ensure its safe application. KFS-EC was relatively stable (~1 log decrease) under stressed conditions such as temperatures (20 °C–50 °C), pHs (3–11), organic solvents (ethanol and chloroform), and biocides (0.1% citric acid, 1% citric acid, and 0.1% peracetic acid). KFS-EC was able to inhibit E. coli O157:H7 efficiently at a multiplicity of infection (MOI) of 0.01 for 8 h with greater inhibitory effect and durability and was stable at 4 °C and 22 °C over a 12-week storage period.

Conclusions

Our results suggest that KFS-EC could be used as a biocontrol agent to E. coli O157:H7.


中文翻译:

一种新型大肠杆菌 O157:H7 特异性噬菌体作为生物防治剂的分离和表征。

目的

大肠杆菌O157:H7 是全球公众关注的主要食源性病原体之一。噬菌体(噬菌体)已成为一种有前途的抗生素替代品,用于控制病原菌。在这里,分离、鉴定和表征裂解的大肠杆菌O157:H7 特异性噬菌体 (KFS-EC),以评估其作为大肠杆菌O157:H7 生物防治剂的潜力。

方法

KFS-EC 是从韩国的屠宰场中分离出来的。进行了形态学分析、基因组分析和几个生理测试来识别和表征 KFS-EC。

结果

特异性测试表明,KFS-EC对 60 种测试菌株中的大肠杆菌O157:H7 菌株具有严格的特异性。形态学和系统发育分析证实 KFS-EC 属于Rb49 病毒属、Tevenvirinae亚科和Myoviridae噬菌体家族。KFS-EC基因组由164,725 bp和总共270个编码序列特征组成,其中114个开放阅读框(ORFs)被鉴定为噬菌体功能基因。KFS-EC 不含编码溶原性和致病性的基因,确保其安全应用。KFS-EC 在温度 (20 °C–50 °C)、pH (3–11)、有机溶剂(乙醇和氯仿)和杀菌剂(0.1% 柠檬酸)等压力条件下相对稳定(约 1 log 下降) 、1% 柠檬酸和 0.1% 过乙酸)。KFS-EC 能够在 0.01 的感染复数 (MOI) 下有效抑制大肠杆菌O157:H7 8 小时,具有更大的抑制效果和持久性,并且在 4°C 和 22°C 下储存 12 周后稳定时期。

结论

我们的结果表明,KFS-EC 可用作大肠杆菌O157:H7 的生物防治剂。
更新日期:2020-02-06
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