| 多重耐药大肠杆菌中前噬菌体的分布特征及诱导分离 |
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| 引用本文： | 刘教,刘畅,陈进,王勉之,熊文广,曾振灵.多重耐药大肠杆菌中前噬菌体的分布特征及诱导分离[J].中国农业科学,2022,55(7):1469-1478. |
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| 作者姓名： | 刘教 刘畅 陈进 王勉之 熊文广 曾振灵 |
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| 作者单位： | 华南农业大学/广东省兽药研制与安全评价重点实验室/国家兽医微生物耐药性风险评估实验室,广州 510642 |
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| 基金项目： | 国家自然科学基金面上项目（31872524）； |
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| 摘 要： | 目的]通过调查前噬菌体在多重耐药大肠杆菌中的分布特征、诱导分离以及前噬菌体中耐药基因与毒力基因的流行状况,为研究前噬菌体介导耐药基因在细菌的传播提供科学依据.方法]挑选前期保存的2018-2019年广东省分离的131株禽源多重耐药大肠杆菌进行核酸提取及全基因组测序,将二代测序的结果组装拼接成全基因组序列,上传至噬菌...
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| 关 键 词： | 前噬菌体 大肠杆菌 诱导 分布特征 耐药基因 |
| 收稿时间： | 2021-02-06 |
Distribution Characteristics of Prophage in Multidrug Resistant Escherichia coli as well as Its Induction and Isolation |
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| LIU Jiao,LIU Chang,CHEN Jin,WANG MianZhi,XIONG WenGuang,ZENG ZhenLing.Distribution Characteristics of Prophage in Multidrug Resistant Escherichia coli as well as Its Induction and Isolation[J].Scientia Agricultura Sinica,2022,55(7):1469-1478. |
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| Authors: | LIU Jiao LIU Chang CHEN Jin WANG MianZhi XIONG WenGuang ZENG ZhenLing |
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| Institution: | South China Agriculture University/Guangdong Provincial Key Laboratory of Veterinary Pharmaceutics Development and Safety Evaluation/National Risk Assessment Laboratory for Antimicrobial Resistance of Animal Original Bacteria, Guangzhou 510642 |
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| Abstract: | 【Objective】 This study investigated the distribution characteristics of prophage in multi-drug resistant Escherichia coli, induction and isolation, as well as the prevalence of drug resistance and virulence genes in prophage, so as to provide a scientific basis for the study of prophage-mediated resistance genes in the spread of bacteria. 【Method】 131 multi-drug resistant E. coli isolating from poultry origin in Guangdong Province from 2018 to 2019 were selected in the laboratory for nucleic acid extraction and whole-genome sequencing. The results of second-generation sequencing were assembled and spliced into a whole-genome sequence and uploaded to the phage. The PHASTER network database was compared and analyzed with the existing phage genome sequences in the database. Drug resistance genes and virulence genes were compared on the CGE database, and then the distribution of drug resistance genes and virulence genes on the prophage were obtained. The mild phage was induced by mitomycin C, separated and purified by using the double-layer plate method. 【Result】 The results of the drug sensitivity test of 131 strains of Escherichia coli showed that the drug resistance rates of ampicillin, tetracycline, florfenicol and compound trimethoprim were all more than 90%, followed by cephalosporin antibiotics, gentamicin, ciprofloxacin, meropenem and colistin with all around 50%, and the resistance rate of tigecycline reached 0.2%. All strains showed multi-drug resistance, and they were all multi-drug resistant Escherichia coli. A total of 736 prophage fragments were detected in 131 strains of multi-drug resistant E. coli, including 329 complete prophage, 66 suspicious phages and 341 incomplete phage, which matched with 40, 20 and 52 known database phage species in different percentages, respectively. The gene sequence of the complete prophage showed that it matched the known phage species better, and the sequence similarity was the highest, with an average of 58.53%. The average number of prophages in 131 strains of E. coli was 5.6, and the average total content was 152.4 kb. Prophage genome accounted for 0.58% to 5.87% of its host genome, with 3.0% being the dominant. The length of the prophage genome ranged from 2.8 to 107.9 kb, and the 13.0 kb prophage had the highest frequency, accounting for 9.1% of all prophages. CGE comparison results showed that the genomes of 131 strains of multi-drug-resistant E. coli detected resistance genes mdf (A), lnu (G) and mcr-1 in 18 prophage sequences. The detected numbers of mdf (A), lnu (G) and mcr-1 were 16, 1, and 1, respectively. 71 strains of multi-drug resistant E. coli prophage carried 6 different virulence genes, and some strains carried 2 or 3 virulence genes. There were 62 prophages carrying the telomerase RNA gene terC, 16 prophages carrying the serum survival increasing gene iss, and the outer membrane protease ompT, among which the adhesin gene iha, the cvaC gene and the ABC transporter gene mchF were at 2, 2, 1, and 1, respectively. Mcr-a gene were detected in prophage of 1 strain multi-drug resistant Escherichia coli. The mdf (A) gene and terC gene were the most common resistance genes and virulence genes in prophage, respectively. The results of mild phage induction experiments showed that the success rate of prophage induction was 84.0%, but the probability of plaque appearance was still relatively low. 【Conclusion】Prophages were widely distributed in multi-drug resistant E. coli and carried a variety of resistance genes and virulence genes. Mild phages had a high induction rate, and have the risk of horizontal transmission of resistance genes and virulence genes, and need to be strengthened and sustained monitor. |
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| Keywords: | prophage Escherichia coli induction distribution characteristics resistance genes |
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